PEPTIDES AND METHODS FOR INDUCING CELL DEATH

Abstract

The invention provide isolated peptides, protides and conjugates having novel peptide sequences which are able to induce antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity. The invention also provides a method of inducing programmed cell death in a cell by contacting the cell with an isolated peptide, protide or conjugate described herein. In some aspects, the method can be used in the diagnosis, prevention, or treatment of a disease, such as an infection, cancer, autoimmune disease, or inflammatory disease.

Description

[0001] This application is a continuation of United States Non-provisional application Ser. No. 12/932,298, filed Feb. 22, 2011, which claims the benefit of priority of U.S. Provisional application Ser. No. 61/338,747, filed Feb. 22, 2010, the entire contents of which are incorporated herein by reference.

FIELD OF THE INVENTION

[0002] The present invention relates generally to compositions and methods for inducing cell death, and more specifically to peptides and compositions having antimicrobial, anti-cancer, anti-inflammatory and/or anti-proliferative activity and methods of using the peptides and compositions as therapeutics.

BACKGROUND OF THE INVENTION

[0003] Programmed cell death pathways are known to exist in most if not all organisms on Earth, ranging from microbes to man. Proteins that effect this function, also known as apoptosis, have been identified in human, other mammals, plants, protozoa, fungi, and bacteria, among other forms of life. In humans, these proteins target the mitochondria, causing permeabilization, dissipation of the membrane potential, activation of intracellular signaling pathways, and ultimate death of the cell. Eukaryotic pathogens also contain mitochondria, and mitochondria are now widely accepted by evolutionary biologists to be decedents of specialized symbiotic bacteria in eukaryotic cells.

[0004] Given these close parallels between mitochondria and bacteria, it is contemplated that specific human, eukaryotic or prokaryotic proteins have necessarily evolved to control prokaryotic symbionts (eg. mitochondria, chloroplasts) or competitors, and directly or indirectly prompt death of microbes or infected or abnormal cells. These types of proteins exhibit similarities in structures (eg., cationic helical domains) and mechanisms of action (eg., membrane interaction or perturbation that can lead to programmed cell death). Thus, such proteins may serve as excellent templates for novel therapeutic molecules, and reveal new insights into host-pathogen co-evolution, cancer biology, and other disease prevention, pathogenesis and treatment.

SUMMARY OF INVENTION

[0005] Embodiments of the invention provide isolated peptides, protides and conjugates having novel peptide sequences which are able to induce antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity. Peptides, protides and conjugates provided by the invention comprise, consist essentially of, or consist of amino acid sequences represented by SEQ ID NOS:1-263 and 288-289. In some aspects, peptides, protides and conjugates described herein have conservative amino acid substitutions or alternative residues at specific locations within a peptide sequence. Non-limiting examples of such substitutions or alternative residues include when the amino acid residue is represented by (x) a serine, a threonine, a tryptophan, a H-bond donor residue or a H-bond acceptor residue can be substituted, or alternatively, when the amino acid residue is represented by (b) a lysine, an arginine, an asparagine, a glutamine or a basic residue can be substituted, or alternatively, when the amino acid residue is represented by (j) a cysteine or a thiol residue can be substituted, or alternatively, when the amino acid residue is represented by (o) an anthrylalanine or other non-natural amino acid can be substituted.

[0006] Embodiments of the invention also provide methods of inducing programmed cell death in a cell by exposing the cell to an isolated peptide, protide or conjugate described herein. In some aspects, the methods can be used in the diagnosis, prevention, or treatment of a disease, such as an infection, cancer, autoimmune disease, or inflammatory disease.

BRIEF DESCRIPTION OF THE DRAWINGS

[0007] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0008] FIG. 1 shows the amino acid sequence of exemplary programmed cell death/holin-like proteins (Dnm-1 (SEQ ID NO:280), Bax (SEQ IN NO:281) and Bcl-2 (SEQ ID NO:282)) identified in Homo sapiens.

[0009] FIG. 2 shows the amino acid sequence of exemplary programmed cell death proteins (CidA (SEQ ID NO:283) and LrgA (SEQ IN NO:284)) identified in Staphylococcus aureus.

[0010] FIG. 3 shows the amino acid sequence of exemplary candidate proteins (Perforin 1 from Bos taurus (SEQ ID NO:285), Bcl-2 from Homo sapiens (SEQ ID NO:286), and BCL-W from Homo sapiens (SEQ ID NO:287)) used for the iterative primary structure analysis of the protein databases (Blastp and/or equivalent thereof) available from the National Center for Biotechnology Information utilizing the basic local alignment sequence tool (BLAST).

[0011] FIG. 4 shows an exemplary data score table sorted by alignment score using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0012] FIG. 5 shows exemplary multisequence alignments using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0013] FIG. 6 shows a phylogram of candidate programmed cell death effector peptides using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0014] FIG. 7 shows a similarity alignment of helical region 1 (amino acids-˜450-490) between candidate peptides identified in the phylogram of FIG. 6 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0015] FIG. 8 shows a similarity alignment of helical region 2 (amino acids-˜540-560) between candidate peptides identified in the phylogram of FIG. 6 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0016] FIG. 9 shows a similarity alignment of helical region 3 (amino acids-˜590-620) between candidate peptides identified in the phylogram of FIG. 6 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0017] FIG. 10 shows a cladogram of candidate programmed cell death effector molecule subset 1 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0018] FIG. 11 shows a similarity alignment of helical region 1/subset 1 (amino acids-˜180-225) between candidate programmed cell death effector molecules identified in the cladogram of FIG. 10 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0019] FIG. 12 shows a similarity alignment of helical region 2/subset 1 (amino acids-˜270-290) between candidate programmed cell death effector molecules identified in the cladogram of FIG. 10 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0020] FIG. 13 shows a similarity alignment of helical region 3/subset 1 (amino acids-˜-320-360) between candidate programmed cell death effector molecules identified in the cladogram of FIG. 10 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0021] FIG. 14 shows a similarity alignment of helical region 4/subset 1 (amino acids-˜490-530) between candidate programmed cell death effector molecules identified in the cladogram of FIG. 10 using the multisequence alignment tool ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI.

[0022] FIG. 15 shows a secondary structure diagram of human Bcl-2, isoform 1 or 2. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence NREIVMKYIHYKLS (residues 1-14 of SEQ ID NO:48) of a peptide predicted to have antimicrobial activity is shown in yellow.

[0023] FIG. 16 shows a secondary structure diagram of human Bcl-2, isoform 1 or 2. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence HLALRQAGDDFSRRYR having a peptide predicted to have antimicrobial activity (SEQ ID NO:53) is shown in yellow.

[0024] FIG. 17 shows a secondary structure diagram of human Bcl-xL. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence SQSNRELVVDFLSYKLSQK (SEQ ID NO:288) of a peptide predicted to have antimicrobial activity is shown in yellow. The amino acid sequence SQSNRELVVDFLSYKLSQK (SEQ ID NO:288) has also been identified as being conserved in multiple PCD-effector templates, including human Bcl-xL and Bcl-xβ, murine Bcl-xγ, and various related proteins.

[0025] FIG. 18 shows a secondary structure diagram of human Bcl-W. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence TRALVADFVGYKLRQK (residues 1-16 of SEQ ID NO:14) of a peptide predicted to have antimicrobial activity is shown in yellow.

[0026] FIG. 19 shows a secondary structure diagram of human Bax. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence RVVALFYFASKLVLKALCTK (residues 1-20 of SEQ ID NO:7) of a peptide predicted to have antimicrobial activity is shown in yellow.

[0027] FIG. 20 shows a secondary structure diagram of human CTL Granulysin. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence RDYRTCLTIVQKLKKM having a peptide predicted of have antimicrobial activity (residues 3-17 of SEQ ID NO:224) is shown in yellow.

[0028] FIG. 21 shows secondary structure diagram of human CTL Granulysin. Cylinders represent alpha helices. The colors are green for alpha helices, orange for beta strands, and blue for coils. The arrows on the helix cylinders point in the N-terminal to C-terminal direction. The amino acid sequence QKLKKIVIVDKPTQRSVSN (SEQ ID NO:289) of a peptide predicted to have antimicrobial activity is shown in yellow.

[0029] FIG. 22 shows a secondary structure ribbon diagram of human Bax (1F16) protein. Helix-1, residues 104-129 are represented in red.

[0030] FIG. 23 shows a secondary structure ribbon diagram of human Bax Helix-1, residues 104-129. Structure A shows the location of positive residues in blue. Structure B shows the most hydrophilic residues in blue and the most hydrophobic in brown.

[0031] FIG. 24 shows a secondary structure ribbon diagram of human Bax (1F16) protein. Helix-2, residues 168-190 are represented in red.

[0032] FIG. 25 shows a secondary structure ribbon diagram of human Bax Helix-2, residues 168-190. Structure A shows the location of positive residues in blue. Structure B shows the most hydrophilic residues as represented in blue and the most hydrophobic residues as represented in brown.

[0033] FIG. 26, panels A-D, show three dimensional alignments between human Bax Helix-1, residues 104-129 vs. IL-8 helix, residues 55-72. For the alignment analyses, comparative sequences of X and Y length are entered, and the computation prioritizes which span of those length are most comparable. Panel A shows a sequence alignment based on the following structural alignment. Panel B shows a horizontal view of a ribbon diagram alignment between Bax, residues 104-129 (Red) and IL-8, residues 56-72 (Blue), whereas panel C shows an axial view of the same alignment. Panel D shows the same ribbon alignment as panel B, wherein the most hydrophilic residues are represented in blue and the most hydrophobic residues are represented in brown. Also included in the figure are the root mean square deviation (RMSD) score and other results from the alignment.

[0034] FIG. 27, panels A-D, show three dimensional alignments between human Bax helix, residues 168-190 vs. IL-8 helix, residues 55-72. For the alignment analyses, comparative sequences of X and Y length are entered, and the computation prioritizes which span of those length are most comparable. Panel A shows a sequence alignment based on the following structural alignment. Panel B shows a horizontal view of a ribbon diagram alignment between Bax, residues 168-190 (Red) and IL-8, residues 55-72 (Blue), whereas panel C shows an axial view of the same alignment. Panel D shows the same ribbon alignment as panel B, wherein the most hydrophilic residues are represented in blue and the most hydrophobic residues are represented in brown. Also included in the figure are the root mean square deviation (RMSD) score and other results from the alignment.

[0035] FIG. 28, panels A-D, show three dimensional alignments between human Bax helix, residues 104-129 vs. magainin residues 1-16. For the alignment analyses, comparative sequences of X and Y length are entered, and the computation prioritizes which span of those length are most comparable. Panel A shows a sequence alignment based on the following structural alignment. Panel B shows a horizontal view of a ribbon diagram alignment between Bax, residues 104-129 (Red) and magainin, residues 1-16 (Blue), whereas panel C shows an axial view of the same alignment. Panel D shows the same ribbon alignment as panel B, wherein the most hydrophilic residues are represented in blue and the most hydrophobic residues are represented in brown. Also included in the figure are the root mean square deviation (RMSD) score and other results from the alignment.

[0036] FIG. 29, panels A-D, show three dimensional alignments between human Bax helix, residues 168-190 vs. magainin residues 1-16. For the alignment analyses, comparative sequences of X and Y length are entered, and the computation prioritizes which span of those length are most comparable. Panel A shows a sequence alignment based on the following structural alignment. Panel B shows a horizontal view of a ribbon diagram alignment between Bax, residues 168-190 (Red) and magainin, residues 1-16 (Blue), whereas panel C shows an axial view of the same alignment. Panel D shows the same ribbon alignment as panel B, wherein the most hydrophilic residues are represented in blue and the most hydrophobic residues are represented in brown. Also included in the figure are the root mean square deviation (RMSD) score and other results from the alignment.

[0037] FIG. 30 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4, LrgA-I-4 and CidA-II-12 against pathogenic bacteria and fungi at pH 7.5. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar. See Tables 22 and 23 for peptide and microorganism designations.

[0038] FIG. 31 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4, LrgA-I-4 and CidA-II-12 against pathogenic bacteria and fungi at pH 5.5. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar. See Tables 22 and 23 for peptide and microorganism designations.

[0039] FIG. 32 shows a histogram of the antimicrobial spectra of exemplary peptides Dnm2-II-4, Dnm1-IV-2, Ncl-VIII-6 and Mfn1-II-2 against pathogenic bacteria and fungi at pH 7.5. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar. See Tables 22 and 23 for peptide and microorganism designations.

[0040] FIG. 33 shows a histogram of the antimicrobial spectra of exemplary peptides Dnm2-II-4, Dnm1-IV-2, Ncl-VIII-6 and Mfn1-II-2 against pathogenic bacteria and fungi at pH 5.5. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar. See Tables 22 and 23 for peptide and microorganism designations.

[0041] FIG. 34 shows a histogram of the antimicrobial spectra of exemplary peptides BclWP-I-4, Csp3-II-12, BclXb-I-2 and BaxP-I-18 against pathogenic bacteria and fungi at pH 7.5. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar. See Tables 22 and 23 for peptide and microorganism designations.

[0042] FIG. 35 shows a histogram of the antimicrobial spectra of exemplary peptides BclWP-I-4, Csp3-II-12, BclXb-I-2 and BaxP-I-18 against pathogenic bacteria and fungi at pH 5.5. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar. See Tables 22 and 23 for peptide and microorganism designations.

[0043] FIG. 36 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Pseudomonas aeruginosa CRM27853 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0044] FIG. 37 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Pseudomonas aeruginosa PA 01 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0045] FIG. 38 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Pseudomonas aeruginosa XEN 5 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0046] FIG. 39 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii 19606 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0047] FIG. 40 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii 17978 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0048] FIG. 41 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter calcoaceticus 23055 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0049] FIG. 42 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter haemolyticus 17906 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0050] FIG. 43 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #ATCC at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0051] FIG. 44 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #1 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0052] FIG. 45 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #6 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0053] FIG. 46 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #12 at pH 5.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0054] FIG. 47 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Pseudomonas aeruginosa CRM27853 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0055] FIG. 48 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Pseudomonas aeruginosa PA 01 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0056] FIG. 49 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Pseudomonas aeruginosa XEN 5 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0057] FIG. 50 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii 19606 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0058] FIG. 51 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii 17978 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0059] FIG. 52 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter calcoaceticus 23055 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0060] FIG. 53 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter haemolyticus 17906 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0061] FIG. 54 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #ATCC at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0062] FIG. 55 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #1 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0063] FIG. 56 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #6 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

[0064] FIG. 57 shows a histogram of the antimicrobial spectra of exemplary peptides Hol-III-4 (SEQ ID NO. 268) and NCl-VIII-6 (SEQ ID NO. 152) against Acinetobacter baumannii HUMC #12 at pH 7.5. Positive control peptides RP1, 6W-RP1, IK and PMP-2, whereas the negative control of double-distilled water (DDH2O) are also shown. The size of the complete zone of inhibition (ZOI) is represented by a blue bar, whereas the size of the partial (ZOI) is represented by a red bar.

DETAILED DESCRIPTION OF THE INVENTION

[0065] Over the last several years, unforeseen structural, functional, and evolutionary relationships among host defenses and other proteins across all kingdoms of life have been discovered. In the course of the studies described herein, unifying themes among polypeptides based on sequence formulae, functional mechanisms, and/or 3D structures have been identified. Embodiments provided by the invention are based in part on the observations that 1) programmed cell death and apoptosis pathway proteins contain archetype sequences that confer membrane interacting/modifying domains similar to those of antimicrobial or other host defense peptides; 2) such sequences encode helical or other sequence and/or 3D structural signatures; and 3) such peptides exert antimicrobial and anti-cancer cell activities. Without being bound by theory, such peptides induce or regulate programmed cell death or related responses in target cells (e.g. microbial pathogens, cancer cells, etc.) leading to death of these cells. It is also contemplated that the mechanisms of action of peptides based on programmed cell death may activate archetypal apoptosis pathways in target cells, thus killing the target cell and circumventing resistance to existing antimicrobial, anti-cancer, or other preventive or therapeutic agents.

[0066] Embodiments of the invention provide that novel antimicrobial, anti-cancer, anti-inflammatory and/or anti-proliferative activity peptides reside in peptide sequences of programmed cell death effector proteins by virtue of the evolutionary necessity for control of microbial and cancer cell survival by increasingly complex eukaryotic systems/symbionts. Thus mitochondrial, chloroplast, and/or nuclear-encoded proteins capable of activating and/or modulating programmed cell death pathways are contemplated to be evolutionary relatives/descendents of polypeptides that originally provided a survival advantage in the face of microbial or neoplastic challenge.

[0067] The peptides, protides and conjugates described herein have the potential to create, augment, or improve several existing therapeutic, prophylactic, diagnostic, and basic research problems. For example, therapeutically, these peptides, protides and conjugates can address the problem of antibiotic-resistant infections and antineoplastic-resistant cancers. Likewise, the peptides, protides and conjugates may serve as immunotherapeutic agents to enhance or restore efficacy of endogenous host defenses. As adjunctive agents, these peptides, protides and conjugates will increase efficacy of conventional agents (such as antibiotics or anti-neoplastic agents), enhance immune functions, and activate or inactivate apoptotic mechanisms of cell regulation associated with aging or other degenerative conditions, and many other potential applications. The scope and diversity of other uses for these peptides, protides and conjugates are considerable. For example, the peptides, protides and conjugates described herein can be used as diagnostic probes in isotopic or non-isotopic forms to localize or characterize diseases or conditions containing signatures such as those characteristic of microbial, neoplastic, necrotic, apoptotic, or other tissues or cells. Additionally, extensions of the above concepts are applicable to the construction, design, delivery, and use of such peptides as research reagents.

[0068] As will be clear to those skilled in the art, the above novel concepts relating to structure-activity relationships in programmed cell death proteins enabled the design of novel antimicrobial, anti-cancer, anti-inflammatory and anti-proliferative peptides, protides and conjugates. These peptides and compositions are useful as diagnostic, prophylactic, and/or therapeutic agents that exploit programmed cell death pathways in pathogens, cancer cells, autoimmune cells, and other disease-caused cells and tissues. Specific examples of peptides, variants, congeners, and mimetics of these molecules are included herein. Embodiments of the invention provide conjugates in which one given molecule can represent or include one or more antimicrobial, anti-cancer, anti-inflammatory, immunomodulatory peptide and one or more non-peptide functional motifs or domains, or combinations of these. Embodiments of the invention also provide protides which are multifunctional and context-activated polypeptides that have two or more effectors with individually distinct biological functions and one or more corresponding activator sites that can each initiate or amplify the biological function of one or more effectors upon context-activation. Therefore, peptides, protides and conjugates exemplified herein are relevant to Antibiotide, Immodulotide, Antineotide, Apoptide, and/or Cascatide class peptides.

[0069] The novel concepts, peptide design strategies, and exemplifying peptides encompass conceptual as well as material inventions. Moreover, variations upon these fundamental themes are applicable to novel therapeutic agents and strategies in virtually any area of medicine, including, but not limited to diagnosis, prevention, and therapy of infectious diseases, cancer and cancer-like diseases, immune and autoimmune disorders, cardiology, aging, and/or other conditions or disease states. Furthermore, the novel peptides based on programmed cell death effectors described herein represent agents and strategies to treat human, animal, and agricultural diseases. Other applications include their use in diagnosis, prevention, or research of diseases, or as research tools to investigate pathogenesis, apoptosis, or related biological phenomena.

[0070] Embodiments of the invention, herein provide an isolated peptide comprising one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 3, 4, 6, 8, 10, 11, 13, 17, 18, 19, 21-25, 30, 31-36, 39-47, 49-52, 54-57, 59-63, 66-75, 84-93, 102-106, 108-121, 132-175, 179-187, 191-199, 205-209, 211-223, 227-235, 238-243, 245-247, 249-251, 253-256 and 260-263, wherein the amino acid residue represented by (x) is a serine, a threonine, a tryptophan, a H-bond donor residue or a H-bond acceptor residue, wherein the amino acid residue represented by (b) is a lysine, an arginine, an asparagine, a glutamine or a basic residue, wherein the amino acid residue represented by (j) is a cysteine or a thiol residue, wherein in the amino acid residue represented by (o) is an anthrylalanine or other non-natural amino acid and wherein the peptide induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity.

[0071] In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bax protein, which are represented by amino acid sequences of SEQ ID NOS: 3, 4, 6, 8, 10, 11, 13, 264, 270 and 271. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bcl-W protein, which are represented by amino acid sequences of SEQ ID NOS: 17, 18, 19, 21-25, 269 and 272. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bcl-xβ protein, which are represented by amino acid sequences of SEQ ID NOS: 30, 31-36 and 273. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bak protein, which are represented by amino acid sequences of SEQ ID NOS: 39-47. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bcl-2 protein, which are represented by amino acid sequences of SEQ ID NOS: 49-52. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bcl-2 isoform 1 protein, which are represented by amino acid sequences of SEQ ID NOS: 54-57. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Mth-1 protein, which are represented by amino acid sequences of SEQ ID NOS: 59-63 and 274. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Mfn-2 protein, which are represented by amino acid sequences of SEQ ID NOS: 66-75. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Dnm-1 protein, which are represented by amino acid sequences of SEQ ID NOS: 84-93 and 275. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Dnm-2 protein, which are represented by amino acid sequences of SEQ ID NOS: 102-106, 108-121, 267, 276 and 277. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Ncl protein, which are represented by amino acid sequences of SEQ ID NOS: 132-175. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Csp3 protein, which are represented by amino acid sequences of SEQ ID NOS: 179-187, 266 and 278. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Bad protein, which are represented by amino acid sequences of SEQ ID NOS: 191-199. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Prf-1 protein, which are represented by amino acid sequences of SEQ ID NOS: 205-209 and 211-223. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Granulysin protein, which are represented by amino acid sequences of SEQ ID NOS: 227-235. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the CidA protein, which are represented by amino acid sequences of SEQ ID NOS: 238-243, 245-247, 265 and 279. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the LrgA protein, which are represented by amino acid sequences of SEQ ID NOS: 249-251. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Lambda S21 protein, which are represented by amino acid sequences of SEQ ID NOS: 253-256. In one aspect, the isolated peptide comprises one or more amino acid sequence, identified from the Holin protein, which are represented by amino acid sequences of SEQ ID NOS: 260-263 and 268.

[0072] Embodiments of the invention provide an isolated peptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NOS: 1, 2, 5, 7, 9, 12, 14-16, 20, 26-29, 37, 38, 48, 53, 58, 64, 65, 72, 76-83, 94-101, 107, 114, 122-131, 170, 176-178, 188-190, 200-204, 210, 224-226, 236, 237, 244, 248, 252, 257-259 and 288-289, wherein the peptide induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity.

[0073] In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Bax protein, which are represented by amino acid sequences of SEQ ID NOS: 1, 2, 5, 7, 9 and 12. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Bcl-W protein, which are represented by amino acid sequences of SEQ ID NOS: 14-16 and 20. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Bcl-xβ protein, which are represented by amino acid sequences of SEQ ID NOS: 26-29. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Bak protein, which are represented by amino acid sequences of SEQ ID NOS: 37 or 38. In one aspect, the isolated peptide consists of the amino acid sequence, identified from the Bcl-2 protein, which is represented by amino acid sequence of SEQ ID NO: 48. In one aspect, the isolated peptide consists of the amino acid sequence, identified from the Bcl-2 isoform 1 protein, which is represented by amino acid sequence of SEQ ID NOS: 53. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Mfn-1 protein, which are represented by amino acid sequences of SEQ ID NOS: 58 or 64. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Mfn-2 protein, which are represented by amino acid sequences of SEQ ID NOS: 65 or 72. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Dnm-1 protein, which are represented by amino acid sequences of SEQ ID NOS: 76-83. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Dnm-2 protein, which are represented by amino acid sequences of SEQ ID NOS: 94-101, 107 and 114. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Ncl protein, which are represented by amino acid sequences of SEQ ID NOS: 122-131 and 170. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Csp3 protein, which are represented by amino acid sequences of SEQ ID NOS: 176-178. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Bad protein, which are represented by amino acid sequences of SEQ ID NOS: 188-190. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Prf-1 protein, which are represented by amino acid sequences of SEQ ID NOS: 200-204 and 210. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Granulysin protein, which are represented by amino acid sequences of SEQ ID NOS: 224-226. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the CidA protein, which are represented by amino acid sequences of SEQ ID NOS: 236, 237 and 244. In one aspect, the isolated peptide consists of the amino acid sequence, identified from the LrgA protein, which is represented by amino acid sequence of SEQ ID NOS: 248. In one aspect, the isolated peptide consists of the amino acid sequence, identified from the Lambda S21 protein, which is represented by amino acid sequences of SEQ ID NOS: 252. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from the Holin protein, which are represented by amino acid sequences of SEQ ID NOS: 257-259. In one aspect, the isolated peptide consists of one or more amino acid sequence, identified from human Bcl-xL protein, which is represented by the amino acid sequence SEQ ID NO: 288 or human CTL Granulysin, which is represented by the amino acid sequence SEQ ID NO: 289. In one aspect, an isolated peptide as described herein has a C-terminus comprising a carboxamide.

[0074] Embodiments of the invention are intended to be used as in ways similar to antibiotic, anti-cancer, or similar medical administration either as local (e.g. topical, oral rinse, inhaled, nebulized, etc.) or systemic (oral ingestion, intravenous, intramuscular, etc) agents. Additionally, the peptides may be used as research tools for basic molecular biology, microbiology, biochemistry or other disciplines as they relate broadly to cellular or molecular biology, infection and immunity, cell regulation and apoptosis, gene expression, signal transduction, or any other area of investigation in which a concept, approach, or specific peptide or may be used.

[0075] In certain embodiments, the invention provides novel isolated peptides having one or more continuous amino acids sequences. As used herein, a "peptide" generally has from about 3 to about 100 amino acids, whereas a polypeptide or protein has about 100 or more amino acids, up to a full length sequence translated from a gene. Additionally, as used herein a peptide can be a subsequence or a portion of a polypeptide or protein. In certain embodiments the size of at least one peptide may comprise, but is not limited to, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 amino acid residues.

[0076] As used herein, an "amino acid residue" refers to any naturally or non-naturally occurring amino acid, any amino acid derivative or any amino acid mimic known in the art. In certain embodiments, the residues of the peptide are sequential, without any non-amino acid interrupting the sequence of amino acid residues. In other embodiments, the sequence may comprise one or more non-amino acid moieties. In particular embodiments, the sequence of residues of the peptide may be interrupted by one or more non-amino acid moieties. Accordingly, the term peptide encompasses amino acid sequences comprising at least one of the 20 common amino acids found in naturally occurring proteins, or at least one modified or unusual or non-natural amino acid, including, but not limited to, Anthrylalanine, 2 Aminoadipic acid (Aad), N Ethylasparagine (EtAsn), 3 Aminoadipic acid (Baad), Hydroxylysine (Hyl), β alanine, β Amino propionic acid (Bala), allo Hydroxylysine (AHyl), 2 Aminobutyric acid (Abu), 3 Hydroxyproline (3Hyp), 4 Aminobutyric acid (4Abu), 4 Hydroxyproline (4Hyp), 6 Aminocaproic acid (Acp), Isodesmosine (Ide), 2 Aminoheptanoic acid (Ahe), allo Isoleucine (AIle), 2 Aminoisobutyric acid (Aib), N Methylglycine (MeGly), 3 Aminoisobutyric acid (Baib), N Methylisoleucine (MeIle), 2 Aminopimelic acid (Apm), 6 N Methyllysine (MeLys), 2,4 Diaminobutyric acid (Dbu), N Methylvaline (MeVal), Desmosine (Des), Norvaline (Nva), 2,2'Diaminopimelic acid (Dpm), Norleucine (Nle), 2,3 Diaminopropionic acid (Dpr), Ornithine (Orn), or N Ethylglycine (EtGly).

[0077] A peptide containing one or more mimetic structures having a similar charge and spatial or steric arrangements as the reference amino acid residues is included within the definition of the term so long as the peptide containing the mimetic portion exhibits a similar or enhanced activity as compared with the reference peptide. It is thus understood that a peptide described herein includes such mimetics as chemically modified peptides, peptide-like molecules containing non-naturally occurring amino acids, and peptoids, which are peptide-like molecules resulting from oligomeric assembly of N-substituted glycines, with similar or enhanced activity as compared with the reference protide upon which the mimetic is derived or having any other property desired by the user, for example, enhanced biostability (see, for example, Goodman and Ro, Peptidomimetics for Drug Design, in "Burger's Medicinal Chemistry and Drug Discovery" Vol. 1 (ed. M. E. Wolff; John Wiley & Sons 1995), pages 803 861), which is incorporated herein by reference in its entirety. Mimetics also include constrained-structures so as to maintain optimal spacing and charge interactions of the amino acid or of the amino acid functional groups. Those skilled in the art know or can determine what structures constitute functionally equivalent amino acid analogs and amino acid mimetics useful for preparation of a peptide described herein.

[0078] Specific examples of amino acid analogs and mimetics can be found described in, for example, Roberts and Vellaccio, The Peptides: Analysis, Synthesis, Biology, Eds. Gross and Meinhofer, Vol. 5, p. 341, Academic Press, Inc., New York, N.Y. (1983), the entire volume of which is incorporated herein by reference. Other examples include peralkylated amino acids, particularly permethylated amino acids. See, for example, Combinatorial Chemistry, Eds. Wilson and Czarnik, Ch. 11, p. 235, John Wiley & Sons Inc., New York, N.Y. (1997), which is incorporated herein by reference in its entirety. Yet other examples include amino acids whose amide portion and, therefore, the amide backbone of the resulting peptide, has been replaced, for example, by a sugar ring, steroid, benzodiazepine or carbo cycle. See, for example, Burger's Medicinal Chemistry and Drug Discovery, supra, Ch. 15, pp. 619 620, which is incorporated herein by reference in its entirety. Methods for synthesizing peptides, polypeptides, peptidomimetics and proteins are well known in the art (see, for example, U.S. Pat. No. 5,420,109; Bodanzsky, Principles of Peptide Synthesis (1st ed. & 2d rev. ed.), Springer-Verlag, New York, N.Y. (1984 & 1993), see Chapter 7; Stewart and Young, Solid Phase Peptide Synthesis, (2d ed.), Pierce Chemical Co., Rockford, Ill. (1984), each of which is incorporated-herein by reference in its entirety).

[0079] In one aspect, the peptide, protide or conjugate can comprise conservatively substituted sequences or alternative residues at specifically identified positions described herein, for example, residues identified in SEQ ID NOS: 3, 4, 6, 8, 10, 11, 13, 17, 18, 19, 21-25, 30, 31-36, 39-47, 49-52, 54-57, 59-63, 66-75, 84-93, 102-106, 108-121, 132-175, 179-187, 191-199, 205-209, 211-223, 227-235, 238-243, 245-247, 249-251, 253-256 and 260-263. In general, a conservative substitution refers to replacement of a given amino acid residue with a residue having similar physiochemical characteristics. Examples of conservative substitutions include (1) non-polar amino acids (Gly, Ala, Val, Leu, and Ile); (2) polar neutral amino acids (Cys, Met, Ser, Thr, Asn, and Gln); (3) polar acidic amino acids (Asp and Glu); (4) polar basic amino acids (Lys, Arg and His); and (5) aromatic amino acids (Phe, Trp, Tyr, and His). Other such conservative substitutions, for example, include substitutions of entire regions having similar hydrophobicity characteristics or substitution of one H-bond donor/acceptor with another H-bond donor/acceptor. An alternative residue refers to a residue that may not be traditionally considered a conservative substitution, but when substituted at the designated position does not adversely effect the functional characteristics of the peptide.

[0080] Hydrogen bonding (H-bond) is a non-covalent type of bonding between molecules or within them, intermolecularly or intramolecularly, and in the context of the peptides described herein include H-bond between amino acids. The H-bond donor is the molecule that has a hydrogen atom bonded to a highly electronegative, small atom with available valence. For example, H--O, H--N, and H--F bonds are extremely polar and as a result, the electron density is easily withdrawn from the hydrogen atom towards the electronegative atom. The partially positive hydrogen in one molecule attracts to partially negative lone pair of the electronegative atom on the other molecule, i.e. an H-bond acceptor, and thus a H-bond forms as a result of such an interaction.

[0081] A "basic" residue refer to an amino acid residue which has a second basic group, which can be, but is not limited to, an amino group (i.e. lysine), a guanidine group (i.e. arginine), or an imidazole ring (i.e. histidine).

[0082] A "thiol" residue refers to an amino acid residue which has a functional sulfur-hydrogen present in the side chain (i.e. cysteine or methionine). A thiol residue, such as cysteine, can also play an important role in the folding and stability of some peptides and proteins through the formation of disulfide bonds.

[0083] A pathological condition appropriate for treatment with a peptide, protide or conjugate described here can be a symptomatic disease or other abnormal condition or injury of a mammalian cell or tissue. Such pathological conditions include, for example, hyperproliferative and unregulated neoplastic cell growth, degenerative conditions, inflammatory diseases, autoimmune diseases and infectious diseases. Hyperplastic and cancer cells proliferate in an unregulated manner, causing destruction of tissues and organs. Specific examples of hyperplasias include benign prostatic hyperplasia and endometrial hyperplasia.

[0084] Abnormal cellular growth can also result from infectious diseases in which foreign organisms cause excessive growth. For example, human papilloma viruses can cause abnormal growth of tissues. The growth of cells infected by a pathogen is abnormal due to the alteration of the normal condition of a cell resulting from the presence of a foreign organism. Specific examples of infectious diseases include DNA and RNA viral diseases, bacterial diseases, fungal diseases, and protozoal or parasitic diseases. Similarly, the cells mediating autoimmune and inflammatory diseases are aberrantly regulated which results in, for example, the continued proliferation and activation of immune mechanisms with the destruction of tissues and organs. Accordingly, "anti-inflammatory activity" refers to a cellular response to a substance or treatment that reduces inflammation and "anti-proliferative activity" refers to a cellular response to a substance that prevents the proliferation or uncontrolled dividing of cells. Specific examples of autoimmune diseases include, for example, rheumatoid arthritis and systemic lupus erythmatosis. Specific examples of degenerative disease include osteoarthritis and Alzheimer's disease. Similarly, the terms infectious diseases, degenerative diseases, autoimmune diseases and inflammatory diseases are intended to include all classes and types of these pathological conditions. Those skilled in the art will know the various classes and types of proliferative, neoplastic, infectious, autoimmune and inflammatory diseases.

[0085] By specific mention of the above categories of pathological conditions, those skilled in the art will understand that such terms include all classes and types of these pathological conditions. For example, the term cancer is intended to include all known cancers, whether characterized as malignant, benign, soft tissue or solid tumors, or hematologic tumors relating to cells in circulation, such as Anal Cancer, Basal Cell Carcinoma, Bladder Cancer, Bone Cancer (Osteosarcoma and Malignant Fibrous Histiocytoma), Brain Tumor, Breast Cancer, Bronchial Tumors, Burkitt Lymphoma, Central Nervous System Lymphoma, Cervical Cancer, Childhood Cancers, Colon Cancer, Colorectal Cancer, Eye Cancer (Intraocular Melanoma or Retinoblastoma), Gallbladder Cancer, Gastric (Stomach) Cancer, Germ Cell Tumor, Head and Neck Cancer, Kidney (Renal Cell) Cancer, Lip and Oral Cavity Cancer, Liver Cancer, Lung Cancer (Non-Small Cell), Lung Cancer (Small Cell), Neuroblastoma, Oral Cancer (Oropharyngeal Cancer), Ovarian Epithelial Cancer, Pancreatic Cancer, Paranasal Sinus and Nasal Cavity Cancer, Parathyroid Cancer, Penile Cancer, Prostate Cancer, Rhabdomyosarcoma, Salivary Gland Cancer, Sarcoma, Skin Cancer (Nonmelanoma), Skin Cancer (Melanoma), Skin Carcinoma (Merkel Cell), Testicular Cancer, Throat Cancer, Thymoma and Thymic Carcinoma, Thyroid Cancer, Urethral Cancer and Vaginal Cancer. Accordingly, "anti-cancer activity" refers to a cellular response to a substance that kills or inhibits the growth of a cancer cell. Cancer cells typically display uncontrolled growth (division beyond the normal limits), invasion (intrusion on and destruction of adjacent tissues), and sometimes metastasis (spread to other locations in the body via lymph or blood). Peptides, protides and conjugates described herein which have anti-cancer activity can kill the cancer cell or prevent the invasion or metastasis of the cancer cell into other tissues. It is contemplated that the mechanism of action through which this activity occurs is through the programmed cell death or related responses in the cells.

[0086] "Antimicrobial activity" refers to a cellular response to a substance that kills or inhibits the growth of a microorganism, such as bacteria, fungi or protozoans. Peptides described herein which have antimicrobial activity can either kill the microorganism (microbicidal) or prevent the growth of the microorganism (microbistatic). In some aspects, the peptides, protides and conjugates described herein show antimicrobial activity again pathogenic microorganisms. A "pathogenic microorganism" refers to a microorganism that causes a disease, disorder or condition, which is commonly referred to as an infection. Pathogenic microorganisms are well known to one of skill in the art and include pathogenic bacteria such as Acinotobacter baumannii, Acinotobacter calcoaceticus, Acinotobacter haemolyticus, Bordetella pertussis, Borrelia burgdorferi, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campylobacter jejuni, Chlamydia pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium tetani, Corynebacterium diphtheriae, Enterococcus faecalis, Enterococcus faecum, Escherichia coli, Francisella tularensis, Haemophilus influenzae, Helicobacter pylori, Legionella pneumophila, Leptospira interrogans, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycoplasma pneumoniae, Neisseria gonorrhoeae, Neisseria meningitides, Pseudomonas aeruginosa, Rickettsia rickettsii, Salmonella typhi, Salmonella typhimurium, Shigella sonnei, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Treponema pallidum, Vibrio cholerae and Yersinia pestis, pathogenic fungi such as Cryptococcus neoformans, Aspergillus fumigatus, Aspergillus flavus, Aspergillus clavatus, Cryptococcus neoformans, Cryptococcus laurentii, Crytococcus albidus, Histoplasma capsulatum, Pneumocystis jirovecii, Stachybotrys chartarum and several members of the Canidida species, such as C. albicans, C. glabrata, C. tropicalis, C. stellatoidea, C. glabrata, C. Krusei, C. parapsilosis, C. guilliermondii, C. viswanathii and C. lusitaniae. Additional examples of pathogenic microorganisms are described by Jorgensen and Pfaller in "A Clinician's Dictionary of Pathogenic Microorganisms" ASM Press (2004), which is herein incorporated by reference in its entirety.

[0087] Human infections due to antibiotic-resistant bacteria and fungi are increasing in frequency and severity. Microbial pathogens exhibiting resistance to one or more antibiotics can now commonly be found in community and nosocomial settings. Antibiotic resistant pathogens currently of the greatest concern are methicillin (multiple) resistant Staphylococcus aureus (MRSA), vancomycin resistant Enterococcus faecalis and Enterococcus faecium (VRE), multi-drug-resistant Streptococcus pneumoniae (MDRSPn) or Streptococcus pyogenes (MDRBRSPy), Pseudomonas aeruginosa (MDRA), and Candida albicans (MDRCA).

[0088] "Programmed cell death" or "PCD" is death of a cell in any form, mediated by an intracellular program. In contrast to other types of cell death, such as necrosis, PCD is carried out as a regulated process which generally confers advantages during an organism's life-cycle. A strategic advantage of apoptosis is that it typically does not induce broader inflammatory responses, which can be injurious and/or delay wound healing, whereas necrosis often induces considerable inflammation. PCD is commonly categorized by two types, apoptosisin (type I cell-death) or autophagic (type II cell death). Apoptosisin, also known as apoptosis, is a series of biochemical events leading to a characteristic cell morphology including blebbing, loss of membrane asymmetry and attachment, cell shrinkage, nuclear fragmentation, chromatin condensation and chromosomal DNA fragmentation known as laddering. Autophagic PCD is a catabolic process involving the degradation of a cell's own components and organelles through the lysosomal machinery prior to the nucleus being destroyed. Additionally, in some aspects, PCD refers to other pathways that have been described including non-apoptotic (i.e. caspase-independent) programmed cell-death, necrosis-like programmed cell death, anoikis, excitotoxicity and Wallerian degeneration.

[0089] Embodiments of the invention provide a context-activated protide having at least one activator site and two or more effectors, wherein at least one effector comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 3, 4, 6, 8, 10, 11, 13, 17, 18, 19, 21-25, 30, 31-36, 39-47, 49-52, 54-57, 59-63, 66-75, 84-93, 102-106, 108-121, 132-175, 179-187, 191-199, 205-209, 211-223, 227-235, 238-243, 245-247, 249-251, 253-256 and 260-263, wherein the amino acid residue represented by (x) is a serine, a threonine, a tryptophan, a H-bond donor residue or a H-bond acceptor residue, wherein the amino acid residue represented by (b) is a lysine, an arginine, an asparagine, a glutamine or a basic residue, wherein the amino acid residue represented by (j) is a cysteine or a thiol residue, wherein the amino acid residue represented by (o) is an anthrylalanine or other non-natural amino acid and wherein the at least one effector induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity. Embodiments of the invention also provide a context-activated protide comprising at least one activator site and two or more effectors, wherein at least one effector comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 1, 2, 5, 7, 9, 12, 14-16, 20, 26-29, 37, 38, 48, 53, 58, 64, 65, 72, 76-83, 94-101, 107, 114, 122-131, 170, 176-178, 188-190, 200-204, 210, 224-226, 236, 237, 244, 248, 252, 257-259 and 288-289, wherein the at least one effector induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity

[0090] In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bax protein, referenced by the amino acid sequence of SEQ ID NOS: 1-13, 264, 270 and 271. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bcl-W protein, referenced by the amino acid sequence of SEQ ID NOS: 14-25, 269 and 272. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bcl-xβ protein, referenced by the amino acid sequence of SEQ ID NOS: 26-36 and 273. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bak protein, referenced by the amino acid sequence of SEQ ID NOS: 37-47. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bcl-2 protein, referenced by the amino acid sequence of SEQ ID NOS: 48-52. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bcl-2 isoform 1 protein, referenced by the amino acid sequence of SEQ ID NOS: 53-57. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Mfn-1 protein, referenced by the amino acid sequence of SEQ ID NOS: 58-64 and 274. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Mfn-2 protein, referenced by the amino acid sequence of SEQ ID NOS: 65-75. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Dnm-1 protein, referenced by the amino acid sequence of SEQ ID NOS: 76-93 and 275. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Dnm-2 protein, referenced by the amino acid sequence of SEQ ID NOS: 94-121, 267, 276 and 277. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Ncl protein, referenced by the amino acid sequence of SEQ ID NOS: 122-175. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Csp3 protein, referenced by the amino acid sequence of SEQ ID NOS: 176-187, 266 and 278. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Bad protein, referenced by the amino acid sequence of SEQ ID NOS: 188-199. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Prf-1 protein, referenced by the amino acid sequence of SEQ ID NOS: 200-223. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Granulysin protein, referenced by the amino acid sequence of SEQ ID NOS: 224-235. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the CidA protein, referenced by the amino acid sequence of SEQ ID NOS: 236-247, 265 and 279. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the LrgA protein, referenced by the amino acid sequence of SEQ ID NOS: 248-251. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Lambda S21 protein, referenced by the amino acid sequence of SEQ ID NOS: 252-256. In one aspect of the invention, the protide has at least one effector comprising an amino acid sequence, identified from the Holin protein, referenced by the amino acid sequence of SEQ ID NOS: 257-263 and 268. In one aspect, the protide has at least one effector comprising an amino acid sequence, identified from human Bcl-xL protein, which is represented by the amino acid sequence SEQ ID NO: 288 or human CTL Granulysin, which is represented by the amino acid sequence SEQ ID NO: 289.

[0091] In one aspect, the activator site is context-activated. The invention also provides that upon context-activation, the protide initiate programmed cell death of the target cells. In some aspects, the context-activation results from a physiological condition, such as, but not limited to acidity, alkalinity, ionic strength or osmotic strength. In some aspects, the context-activation results from association with an activator molecule. The activator molecules can modify the activator site upon association. In some aspects, modification of the activator site includes cleavage of the activator site. In some aspects, the activoator molecule is an enzyme, such as a protease, esterase or lipase. In one aspect, the activator is expressed by a pathogenic microorganism as described herein. In another aspect, the activator is present in the context of a tumor cell, such as a tumor-specific protease. An example of a tumor-specific protease is a matrix bound protein such as matrix metalloproteinase. In another aspect, the activator is present in the context of an inflammatory response, wherein activators such as thrombin, bradykinin, elastase and metalloproteinase are expressed.

[0092] The term "protide," as used herein, refers to a mosaic molecule composed of two or more peptide or non-peptide functional domains, referred to as effectors, and one or more corresponding activator sites. A protide can consist of an indefinite number of effector and activator domains that can vary in function, activation, position, continuity, or sequence. Additional examples of protide compositions and designs are described in U.S. Patent Application Publication 2006-0074016, 2006-0135416 and U.S. Pat. No. 7,067,621, which are herein incorporated by reference.

[0093] The protides described herein have two or more distinct biological functions and are designed to be activated within a defined or characteristic context. The protides described herein have at least one activator site and two or more effectors, wherein at least one of the effectors has and amino acid sequence of a peptide described herein. Protides have the advantage of designs that can be customized, engineered, chosen, or combined to allow for highly selective correspondence to or association with or unique to a specific pathological condition or etiology. The distinct biological functions can further be associated with distinct functional aims, for example, therapy, prevention, amplification and detoxification. As described herein, a multifunctional, context-activated protide can be designed to be activated in any context desired by the user, a feature which makes the protides useful to applications in many areas of medicine and biomedical research, including, for example, diagnosis, imaging, detection, speciation or other specification, prevention/prophylaxis, and therapy of a wide range of pathological conditions such as infectious diseases, neoplastic diseases, immune and autoimmune disorders, cardiovascular conditions, disorders in metabolism or physiology, diseases of inheritance or genetic abnormality, a variety of pathological conditions associated with gene expression, mitochondrial dysfunction or regulation, as well as cell death and/or cellular senescence.

[0094] As described herein, in addition to their direct antimicrobial efficacies, the peptides, protides and conjugate described herein are useful based on their ability to circumvent or minimize conventional resistance mechanisms by pathogens or tumor cells. For example, this can be the result of activation by activators that are present outside of the target cell such that the peptides, protides or conjugate need not necessarily enter the target cell to be activated and to achieve subsequent efficacy, thus minimizing the likelihood for resistance due to reduced target access or increased efflux of the peptides, protides or conjugate. Furthermore, in many conventional resistance mechanisms, resistance can be induced by the presence of the anti-infective agent itself. In particular, protides can be designed to be activated by such microbial counter-responses or virulence factors. Thus, the more of the activator that is made by the organism, the more protide activation results, yielding an expected amplification of the anti-pathogenic efficacy of the protide. Conversely, decreased production of the activators can translate in turn to decreased presence or function of these same activators such as virulence factors or mediators of pathogenesis, in essence turning off the pathogenic potential of the target cell, or reducing its ability to protect itself from otherwise normal host defenses. Similarly, protides can be beneficial by reconstituting tumor cell or microbial pathogen susceptibility to conventional therapeutic agents, to which these pathogenic cells would otherwise be resistant. Thus, the protides can either be activated from upregulation of resistance- or virulence factor expression, or can impact efficacy by effecting the downregulation of virulence factor expression by pathogenic cells or organisms.

[0095] In applications of the methods described herein, involving an established infection or a host response to infection, activators can be present or generated. An activator useful for activation of a protide of the invention can be advantageously selected based on a high concentration in the immediate proximity of the infection locus so as to allow for activation of the majority of protides in the desired context. One skilled in the art will be able to select an activator that represents the desired activation context. For applications of the invention methods in the arena of microbial infection, context-activation can be designed to specifically occur in the local context of infection so as to effect optimal relative protide effector concentrations in specific contexts of infection. In addition to context activation that maximizes efficacy, the protides and methods of the invention also minimize the potential for inadvertent host cytotoxicity in areas that do not represent the context. Therefore, in the absence of infection, the protide activators are either absent or are present at concentrations insufficient for effective protide activation, thereby minimizing inadvertent or indiscriminant acute toxicity.

[0096] In addition to specific pathogen or host molecules that can serve as activators as described above, protides can also be designed to become activated to diagnose, prevent, or treat infection in unique and/or specific biochemical or physiological contexts associated with microbial pathogens. Examples of such biochemical or environmental contexts include ionic, osmotic, pH, oxidation/reduction, or other conditions that are unique to, characteristic of, or present in the context of infection or disease processes that occur upon infection, or host responses to these events. For example, a protide can be designed to require the influence of protonation, conformation change, or other modification that occurs uniquely or disproportionately in the context of acidic pH, to activate the protide or its ensuing effectors by altering their structure-activity relationship(s) from inactive to active. As one example, genitourinary tissues, such as renal-tissues or genitourinary mucosa, can exhibit pH values that are decreased normally, or in the setting of infection. A protide designed to be activated only under such acidic conditions could be designed to either be vulnerable to activation in these conditions, or directly activated by these conditions, and thus would be predicted to be active only in such contexts. Alternatively, protides can be designed to be inactive in particular contexts or conditions, such as conditions of relatively high osmotic strength or relatively high pH, so as to minimize or prevent untoward or toxic effects such as nephro- or hepatotoxicity. By way of a further example, activation as well as leukocyte accumulation are conditions associated with infection. Moreover, a fundamental strategy of host defense phagocytes is to phagocytize the microbial pathogen, subjecting it to the harsh environment of the acidic phagolysosome. The compartment so created can become acidified to pH values of 5.5 or lower as the leukocyte responds to the pathogen. Therefore, a protide can be designed that is activated or has amplified or antimicrobial activities, for example, by pH, phagolysosomal enzymes or reactants, or a combination of these conditions, or can amplify or potentiate the antimicrobial mechanisms of leukocytes or other host cells within such settings, so as to inhibit or kill pathogens that enter such cells.

[0097] Protide activation also can include conformational, oxidation or reduction-mediated changes in disulfide array, assembly into multimers of two or more homomeric (identical) or heteromeric (non-identical) effectors, or other modifications of the protide and/or its subsequent effectors. In a particular embodiment, protide activation is triggered as a result of protide accumulation, or its resulting effector components, so as to achieve or surpass threshold concentrations required to optimize or catalyze activation or activity through multimerization or other modification in structure or function of the protide or its effectors.

[0098] It is understood, that activation can involve combinations of the protide activation strategies described above. For example, a protide can be designed that is not responsive to an activator unless both the protide and the activator are present within a context associated with or resulting from infection or other disease.

[0099] The term "context-activated," as used herein in reference to a protide of the invention, refers to the initiation, activation or amplification of a biological or other desired, for example, diagnostic or prophylactic function of one or more protide effectors in a particular temporal, spatial, pathological and/or biochemical context. Context-activation can be initiated by direct or indirect interaction between a protide activator site and a corresponding activator that is selectively associated with the particular context. As used herein, context-activation encompasses activation in a wide variety of contexts that can include, for example, local, regional, systemic, and/or temporal proximity; as well as the presence or absence of an etiological agent, pathologic condition, or characteristic components thereof.

[0100] Thus, context need not be limited to a place, time or quality, but also can be the presence or absence of an activator, for example, an enzyme elaborated by an organism such as, for example, a specific strain of bacteria. The context for activation can consequently be of any breadth desired by the user, for example, can target a class of organisms or cell types, for example, by using an activator that is ubiquitous to the targeted class, or can alternatively have a more narrow focus by using an activator that represents a more narrowly defined target, for example, a particular genus, organism, species, subspecies, strain, or cell or tissue type. The context can be associated with a pathological condition, but also can be selected to represent a non-pathological environment, for example, in prophylactic applications of the invention practiced to preserve a normal or homeostatic condition.

[0101] As used herein, the term "effector" refers to the peptide or non-peptide functional domains of a protide provide herein that have specific individual functions, which are initiated or amplified upon activation and achieve specific functions relating to the diagnosis, prevention, or treatment of a disease. As described herein, a protide has at least two effector domains with distinct, complementary and/or synergistic biological functions. An effector is inactive or exhibits relatively reduced or attenuated biological activity unless an activator, by virtue of either its presence or absence, alters or facilitates or allows the altering of its corresponding activator site and, as a result, initiates or amplifies the diagnostic, prophylactic, therapeutic, or other biological function(s) of the effector(s). Multiple effectors can be induced by the same activator site. Peptide and non-peptide effectors can be present in the same protide, which can be referred to as a hybrid protide. Similarly, a protide can consist exclusively of peptide effectors, also referred to as a peptide protide. Similarly, a protide of the invention can consist exclusively of non-peptidic effectors. The biological function(s) of an effector that corresponds to a protide described herein can be, for example, antimicrobial, immunomodulatory, pro- or anti-inflammatory, tumoricidal, pro- or anti-apoptotic, pro- and anti-angiogenic and/or hemolytic.

[0102] As described herein, a protide of the invention can be bifunctional or multifunctional, with two or more unique complementary effectors, and one or more activators as determined by specific effector and activator site domains engineered into the mosaic protide, which can be activated by specific molecules or conditions present in unique or strategic contexts of interest. Examples of such effectors can include one or more antimicrobial, anti-neoplastic, anti-inflammatory, immunomodulatory, or other peptide or non-peptide functional domains, or combinations thereof.

[0103] As used herein, the term "activator site" when used in reference to a protide of the invention, refers to a domain of the protide that, in the presence of an activator, initiates, promotes, amplifies or modulates the specific biological function of one or more effectors. As described herein, an activator site can be modified, cleaved, processed or otherwise altered in the presence of an activator. In addition, an activator site can be sensitive either to the absolute presence or absence of an activator as well as can be sensitive to a threshold concentration of an activator rather than its mere presence.

[0104] An activator site useful in the invention can include one or more sites for cleavage, modification, processing or other triggering by strategic activators, which can be, for example, proteases, esterases, lipases, or other endogenous enzymatic activators or cascades generated by or associated with a specific condition such as, for example, the presence of pathogenic microorganisms, damaged or inflamed tissues, or hematologic or solid neoplastic or pre-neoplastic cells or tumors. Such an activator site also can be selected to exploit contexts associated with biochemical or physical conditions such as requisite acidity or alkalinity, for example, acidic phagolysomes containing intracellular bacteria or fungi; or ionic or osmotic strength, for example, in a renal context, that represent a specific pathologic or non-pathologic context. Furthermore, an activator site can be selected to exploit normal rather than a pathologic context.

[0105] An activator site can be subject to proteolytic as well as non-proteolytic activation. For example, the activator site can be located within the peptide moiety, and require a protease activator. In other embodiments, the non-proteolytic activator can target a non-proteinaceous substrate component of the protide. For example, a protide of the invention can include an esterase activator and can link peptide and/or non-peptide moieties (eg. a protide consisting of peptide and conventional antibiotic effectors) by means of an ester bond. Other biochemically relevant bonds or linkages that can serve as activation sites in an invention protide can include, for example, lipase- (lipid cleaving), nuclease- (nucleic acid cleaving), and kinase or phosphatase- (phosphate addition or removal) sensitive activators that target substrates other than peptides. For example, certain microbial pathogens or tumor cells can express, or abnormally express restriction enzymes that can provide a suitable basis for design of a protide that could be activated only-within the target cell, further reducing indiscriminant host cytotoxicity.

[0106] As used herein, the term "activator" refers to a molecule or condition that, by altering the activator site, causes the liberation or onset of a specific diagnostic or biological function of effector(s). As described herein, an activator can be a normal or abnormal exogenous or endogenous cell, structure or molecule, a condition or milieu (normal or abnormal), or a combination thereof that is associated with a specific context in which activation of the protide is desired. Thus, an activator can be selected based on its presence in a temporal, spatial, or physiological context, which can be normal or abnormal, that is associated with the desired context for protide activation. An activator can consequently include physiological conditions including, for example, acidity, alkalinity, conditions of oxidation or reduction, and/or ionic and/or osmotic strength, that are associated with a particular context, and modulate protide activation. Alternatively, an activator can be a structure or molecule, for example, an enzyme, that is present in a particular spatial, temporal or pathological context. The activator molecule can modify the activator site upon association, for example, by cleavage or other modification that results in activation in the particular context, or can facilitate interaction between protide and activator(s). The activator molecule can be an enzyme including, for example, protease, esterase, lipase, nucleases or peptidase.

[0107] In one embodiment of the invention, an activator site can encompass one or more domains for cleavage, modification, processing or any other type of liberation by an activator, for example, a protease, esterase, lipase or other endogenous or exogenous enzymatic activator or cascade. The choice of one or more activator sites that correspond to specific activators depends directly on the desired context for activation. Thus, an activator can be a particular pathologic setting or condition that is chosen based on its association with a particular etiological agent or host response. In the presence of the activator, one or more effectors are liberated so as to achieve a specific function relating to, for example, the treatment, prevention, or diagnosis of a targeted disease. An activator site can thus be strategically designed to become activated in temporal and spatial proximity to activator expression, thereby allowing the activation of a protide to be targeted to a particular context and over time so as to maximize the desired therapeutic or prophylactic effect, while minimizing untoward or undesirable toxicities or other side effects.

[0108] As described herein, an activator site is selected based on its correspondence and/or association with the context in which the two or more protide effectors are to be liberated so as to initiate or potentiate their functions. Therefore, as long as an activator is associated with the context, the invention can be practiced with any context desired. Those skilled in the art will appreciate that, given the versatility of activators useful for practicing the invention as described herein, a protide can be designed based on virtually any context desired, including, for example, vascular injury, presence of a neoplasm or cancer, infection, and inflammation.

[0109] In one embodiment, the protide is an antimicrobial protide, which also can be referred to as an antimicrotide. Cleavage sites for strategic proteases can be engineered into multifunctional antimicrobial protides so as to represent the activator site of the protide. Upon activation of the protease in the localized or generalized context of tissue injury or infection, as selected by the user, the inactive protide is cleaved, liberating independent and active molecules to effect their respective biological functions. Prior to and beyond the setting of activation of the strategic protease representing the activator, the mosaic protide construct is relatively inactive both with respect to antimicrobial function and host cell toxicity. A mosaic protide construct can consist of an indefinite number (1 through n) of effector and activator domains that can vary in function, activation, position, continuity, or sequence. Effectors corresponding to one or more protides activated by the same or distinct activators also can function synergistically, and/or can recombine in a manner facilitating their complementary functions. As an example, in the context of vascular injury, a protide activator can be selected that specifically represents this particular context, for example, a clotting cascade protease such as thrombin, or a complement fixing protease such as a C3 convertase, for example, C4B2A or C3bBb. Similarly, as another example, a protide activator can be selected that represents a broader constellation of symptoms or conditions, such as sepsis, in which corresponding activators can include serine proteases associated with systemic inflammation, sepsis, or injury, such as activated protein C.

[0110] A further embodiment of the invention encompasses anti-neoplastic protides, which also are referred to as antineotides. Many tumor cells produce or overexpress characteristic activators, such as matrix metalloproteinases (MMP) or other enzymes that are not expressed by, or at levels much higher than normal cells. Consequently, the activator can be a tumor-specific protease, for example, a matrix metalloproteinase or thymidylate synthase (TS), which is overexpressed in the majority of cancers. A tumor-specific protease also can be associated with a more narrow neoplastic context, such as a serine protease that is specifically expressed in prostate cells, for example, PSA, human kallikrein-2 (hK2), human kallikrein-11 (hK11) and TMPRSS2.

[0111] Embodiment of the invention provide a conjugate having one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 3, 4, 6, 8, 10, 11, 13, 17, 18, 19, 21-25, 30, 31-36, 39-47, 49-52, 54-57, 59-63, 66-75, 84-93, 102-106, 108-121, 132-175, 179-187, 191-199, 205-209, 211-223, 227-235, 238-243, 245-247, 249-251, 253-256 and 260-263 and a moiety, wherein the amino acid residue represented by (x) is a serine, a threonine, a tryptophan, a H-bond donor residue or a H-bond acceptor residue, wherein the amino acid residue represented by (b) is a lysine, an arginine, an asparagine, a glutamine or a basic residue, wherein the amino acid residue represented by (j) is a cysteine or a thiol residue, wherein in the amino acid residue represented by (o) is an anthrylalanine or other non-natural amino acid and wherein the conjugate induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity. Embodiments of the invention also provide a conjugate having one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 1, 2, 5, 7, 9, 12, 14-16, 20, 26-29, 37, 38, 48, 53, 58, 64, 65, 72, 76-83, 94-101, 107, 114, 122-131, 170, 176-178, 188-190, 200-204, 210, 224-226, 236, 237, 244, 248, 252, 257-259 and 288-289 and a moiety, wherein said conjugate induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity.

[0112] In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bax protein, referenced by the amino acid sequence of SEQ ID NOS: 1-13, 264, 270 and 271. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bcl-W protein, referenced by the amino acid sequence of SEQ ID NOS: 14-25, 269 and 272. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bcl-xβ protein, referenced by the amino acid sequence of SEQ ID NOS: 26-36 and 273. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bak protein, referenced by the amino acid sequence of SEQ ID NOS: 37-47. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bcl-2 protein, referenced by the amino acid sequence of SEQ ID NOS: 48-52. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bcl-2 isoform 1 protein, referenced by the amino acid sequence of SEQ ID NOS: 53-57. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Mth-1 protein, referenced by the amino acid sequence of SEQ ID NOS: 58-64 and 274. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Mfn-2 protein, referenced by the amino acid sequence of SEQ ID NOS: 65-75. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Dnm-1 protein, referenced by the amino acid sequence of SEQ ID NOS: 76-93 and 275. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Dnm-2 protein, referenced by the amino acid sequence of SEQ ID NOS: 94-121, 267, 276 and 277. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Ncl protein, referenced by the amino acid sequence of SEQ ID NOS: 122-175. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Csp3 protein, referenced by the amino acid sequence of SEQ ID NOS: 176-187, 266 and 278. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Bad protein, referenced by the amino acid sequence of SEQ ID NOS: 188-199. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Prf-1 protein, referenced by the amino acid sequence of SEQ ID NOS: 200-223. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Granulysin protein, referenced by the amino acid sequence of SEQ ID NOS: 224-235. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the CidA protein, referenced by the amino acid sequence of SEQ ID NOS: 236-247, 265 and 279. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the LrgA protein, referenced by the amino acid sequence of SEQ ID NOS: 248-251. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Lambda S21 protein, referenced by the amino acid sequence of SEQ ID NOS: 252-256. In one aspect of the invention, the conjugate has at least one or more amino acid sequence, identified from the Holin protein, referenced by the amino acid sequence of SEQ ID NOS: 257-263 and 268. In one aspect, the conjugate has at least one or more amino acid sequence, identified from human Bcl-xL protein, which is represented by the amino acid sequence SEQ ID NO: 288 or human CTL Granulysin, which is represented by the amino acid sequence SEQ ID NO: 289.

[0113] In one aspect, the moiety comprises a therapeutic agent, a targeting peptide or a label. In one aspect, therapeutic agent is a cytotoxic agent, such as an antibiotic or a chemotherapeutic agent. In one aspect, the targeting peptide selectively homes a conjugate described herein to a microorganism, a tumor tissue, tumor cell or tumor vasculature. In one aspect, the targeting peptide selectively homes the conjugate to an immune regulatory cell or an immune effector cell. In one aspect, the conjugate described herein has a targeting peptide, such as, but not limited to, an antibody or a fragment thereof. In another aspect, the moiety of the conjugate described herein is a label, such as a radioisotope or a dye.

[0114] As used herein, the term "conjugate" refers to a peptide having an amino acid sequence as described herein linked to a moiety. A "moiety" is used broadly to mean a physical, chemical, or biological material that is linked to a peptide for the purpose of targeting the peptide to a select organ, tissue or cell type or providing an additional functional group to the peptide. In particular, a moiety is a biologically useful moiety such as therapeutic moiety, a diagnostic moiety or a drug delivery vehicle. Thus, a moiety can be a therapeutic agent, for example, a cancer chemotherapeutic agent. Cancer chemotherapeutic agents are well known to one of skill in the art and include, without limitation, alkylating agents such as cyclophosphamide, mechlorethamine, chlorambucil and melphalan, anthracyclines such as daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone and valrubicin, cytoskeletal disruptors such as paclitaxel and docetaxel, epothilones such as epothilones A through F, inhibitors of topoisomerase II such as etoposide, teniposide and tafluposide, nucleotide analogs and precursor analogs such as azacitidine, azathioprine, capecitabine, cytarabine, doxifluridine, fluorouracil, gemcitabine, mercaptopurine, methotrexate, and tioguanine, peptide antibiotics, such as bleomycin, platinum-based agents such as carboplatin, cisplatin and oxaliplatin, retinoids such as tretinoin, and vinca alkaloids or their derivatives such as vinblastine, vincristine, vindesine and vinorelbine. Such a moiety when linked to a peptide, provides a conjugate useful for treating a cancer in a subject. In addition, a moiety can be a drug delivery vehicle such as a chambered microdevice, a cell, a liposome or a virus, which can contain an agent such as a drug or a nucleic acid.

[0115] A moiety also can be a targeting peptide or nucleic acid, to which a peptide as described herein is grafted for the purpose of directing the peptide to a selected organ, tissue, tumor or cell (Smith et al., J. Biol. Chem. 269:32788-32795 (1994); Goldman et al., Cancer Res. 15:1447-1451 (1997) and U.S. Pat. No. 6,576,239, each of which is incorporated herein by reference). For example, a targeting peptide or nucleic acid can be expressed as a fusion protein with a desired peptide such that the peptide or nucleic acid targets the grafted peptide to a selected tumor tissue, tumor cell or tumor vasculature. Such a desired peptide, which is grafted to the tumor homing peptide, can be a polypeptide involved in initiating a programmed cell death pathway as described herein or inducing any other cellular response resulting in anti-cancer activity. Additionally, targeting peptides, which can be grafted to a peptide as described herein having antimicrobial activity, include peptides that selectively home to a microorganism. For example, peptide sequences have been identified that selectively bind to surface molecules of fugal pathogens such as invasive Aspergillus species as described in U.S. Patent Application 2005-0187161. Still further, the invention provides a conjugate wherein the targeting peptide selectively homes the desired peptide to a cell involved in the immune response, including immune regulatory cells such as lymphocytes or immune effector cells such as macrophages or granulocytes. Conjugates provided herein include these and other exemplary peptide or nucleic acid sequences grafted to a peptide described herein. tumor tissue, tumor cell, tumor vasculature, immune regulatory cell or immune effector cell.

[0116] A "targeting peptide" is a peptide comprising a contiguous sequence of amino acids, which is characterized by selective localization to an organ, tissue, or cell type. Selective localization may be determined, for example, by methods disclosed below, wherein the putative targeting peptide sequence is incorporated into a protein that is displayed on the outer surface of a phage. Administration to a subject of a library of such phage that have been genetically engineered to express a multitude of such targeting peptides of different amino acid sequence is followed by collection of one or more organs, tissues, or cell types from the subject and identification of phage found in that organ, tissue, or cell type. A phage expressing a targeting peptide sequence is considered to be selectively localized to a tissue or organ if it exhibits greater binding in that tissue or organ compared to a control tissue or organ. Preferably, selective localization of a targeting peptide should result in a two-fold or higher enrichment of the phage in the target organ, tissue, or cell type, compared to a control organ, tissue, or cell type. Selective localization resulting in at least a three-fold, four-fold, five-fold, six-fold, seven-fold, eight-fold, nine-fold, ten-fold or higher enrichment in the target organ compared to a control organ, tissue or cell type is more preferred. Alternatively, a phage expressing a targeting peptide sequence that exhibits selective localization preferably shows an increased enrichment in the target organ compared to a control organ when phage recovered from the target organ are reinjected into a second host for another round of screening. Further enrichment may be exhibited following a third round of screening. Another alternative means to determine selective localization is that phage expressing the putative target peptide preferably exhibit a two-fold, more preferably a three-fold or higher enrichment in the target organ or tissue compared to control phage that express a non-specific peptide or that have not been genetically engineered to express any putative target peptides. Another means to determine selective localization is that localization to the target organ or tissue of phage expressing the target peptide is at least partially blocked by the co-administration of a synthetic peptide containing the target peptide sequence. "Targeting peptide" and "homing peptide" are used synonymously herein.

[0117] A targeting peptide is useful, for example, for targeting a desired peptide to the selected tumor as discussed above. In addition, a targeting peptide in conjunction with a detectable label can be used to identify the delivery of a desired peptide to a sample. As used herein, the term "sample" is used in its broadest sense to mean a cell, tissue, organ or portion thereof, including a tumor, that is isolated from the body. A sample can be, for example, a histologic section or a specimen obtained by biopsy or cells that are placed in or adapted to tissue culture.

[0118] The term "antibody" is well-known in the art and refers to a protein functionally defined as a binding protein and structurally defined as comprising an amino acid sequence that is recognized by one of skill in the art as having variable and constant regions. A typical antibody structural unit is known to comprise a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" and one "heavy" chain. The N-terminal portion of each chain defines the variable region of about 100 to about 110 amino acids, which are primarily responsible for antigen recognition and binding. The terms variable heavy chain (V_H) and variable light chain (V_L) regions refer to these light and heavy chains, respectively. The variable region includes the segments of Framework 1 (FR1), CDR1, Framework 2 (FR2), CDR2, Framework 3, CDR3 and Framework 4 (FR4). Antibodies are typically divided into five major classes, IgM, IgG, IgA, IgD, and IgE, based on their constant region structure and immune function. The constant region is identical in all antibodies of the same isotype, but differs in antibodies of different isotypes. Heavy chains γ, α and δ have a constant region composed of three tandem (in a line) Ig domains, and a hinge region for added flexibility; heavy chains μ and ε have a constant region composed of four immunoglobulin domains. Antibody classes can also be divided into subclasses, for example, there are four IgG subclasses IgG1, IgG2, IgG3 and IgG4. The structural characteristics that distinguish these subclasses from each other are known to those of skill in the art and can include the size of the hinge region and the number and position of the interchain disulfide bonds between the heavy chains. The constant region also determines the mechanism used to destroy the bound antigen. A light chain has two successive regions: one constant region, which are designated as κ and λ, and one variable region.

[0119] As used herein, the term "functional fragment" when used in reference to the antibodies described herein is intended to refer to a portion of the antibody including heavy or light chain polypeptides which still retains some or all or the binding activity of the antibody. Such functional fragments can include, for example, antibody functional fragments such as Fab, F(ab)₂ Fv, and single chain Fv (scFv). Other functional fragments can include, for example, heavy or light chain polypeptides, variable region polypeptides or CDR polypeptides or portions thereof so long as such functional fragments retain binding activity, specificity, inhibitory and activation activity. The term is also intended to include polypeptides encompassing, for example, modified forms of naturally occurring amino acids such as D-stereoisomers, non-naturally occurring amino acids, amino acid analogues and mimetics so long as such polypeptides retain functional activity as defined above.

[0120] A moiety can be a detectable label such a radiolabel or can be a cytotoxic agent, including a toxin such as ricin or a drug such as a chemotherapeutic agent or can be a physical, chemical or biological material such as a liposome, microcapsule, micropump or other chambered microdevice, which can be used, for example, as a drug delivery system. Generally, such microdevices, should be nontoxic and, if desired, biodegradable. Various moieties, including microcapsules, which can contain an agent, and methods for linking a moiety, including a chambered microdevice, to a molecule of the invention are well known in the art and commercially available (see, for example, "Remington's Pharmaceutical Sciences" 18th ed. (Mack Publishing Co. 1990), chapters 89-91; Harlow and Lane, Antibodies: A laboratory manual (Cold Spring Harbor Laboratory Press 1988), each of which is incorporated herein by reference; see, also, Hermanson, Bioconjugate Techniques (Academic Press 1996)).

[0121] A "label" refers a compound has at least one element, isotope or chemical compound attached to enable the detection of the compound. In general, labels fall into three classes: a) isotopic labels, which may be radioactive or heavy isotopes; b) immune labels, which may be antibodies or antigens; and c) colored or fluorescent dyes. The labels may be incorporated into a HIPK1 nucleic acids, proteins and antibodies at any position. For example, the label should be capable of producing, either directly or indirectly, a detectable signal. The detectable moiety may be a radioisotope, such as ³H, ¹⁴C, ³²P, ³5S, or ¹²⁵I, a fluorescent or chemiluminescent compound, such as fluorescein isothiocyanate, rhodamine, or luciferin, or an enzyme, such as alkaline phosphatase, beta-galactosidase or horseradish peroxidase. Any method known in the art for conjugating the antibody to the label may be employed, including those methods described by Hunter et al., Nature, 144:945 (1962); David et al., Biochemistry, 13:1014 (1974); Pain et al., J. Immunol. Meth., 40:219 (1981); and Nygren, J. Histochem. and Cytochem., 30:407 (1982).

[0122] Peptides, protides and conjugates, which are identified herein, can be synthesized in required quantities using routine methods of solid state peptide synthesis or can be purchased from commercial sources (for example, Anaspec; San Jose Calif.) and a desired moiety can be linked to the peptide. Several methods useful for linking a moiety to a peptide are known in the art, depending on the particular chemical characteristics of the molecule. For example, methods of linking haptens to carrier proteins as used routinely in the field of applied immunology (see, for example, Harlow and Lane, supra, 1988; Hermanson, supra, 1996).

[0123] A moiety such as a therapeutic or diagnostic agent can be conjugated to a peptide using, for example, carbodiimide conjugation (Bauminger and Wilchek, Meth. Enzymol. 70:151-159 (1980), which is incorporated herein by reference). Carbodiimides comprise a group of compounds that have the general formula R--N═C═N--R', where R and R' can be aliphatic or aromatic, and are used for synthesis of peptide bonds. The preparative procedure is simple, relatively fast, and is carried out under mild conditions. Carbodiimide compounds attack carboxylic groups to change them into reactive sites for free amino groups. Carbodiimide conjugation has been used to conjugate a variety of compounds to carriers for the production of antibodies.

[0124] The water soluble carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) is particularly useful for conjugating a moiety to a peptide and was used to conjugate doxorubicin to tumor homing peptides (U.S. Patent Application Publication 2004-0131623). The conjugation of doxorubicin and a tumor homing peptide requires the presence of an amino group, which is provided by doxorubicin, and a carboxyl group, which is provided by the peptide.

[0125] In addition to using carbodiimides for the direct formation of peptide bonds, EDC also can be used to prepare active esters such as N-hydroxysuccinimide (NHS) ester. The NHS ester, which binds only to amino groups, then can be used to induce the formation of an amide bond with the single amino group of a moiety. The use of EDC and NHS in combination is commonly used for conjugation in order to increase yield of conjugate formation (Bauminger and Wilchek, supra, 1980).

[0126] Other methods for conjugating a moiety to a peptide can also be used. For example, sodium periodate oxidation followed by reductive alkylation of appropriate reactants can be used, as can glutaraldehyde crosslinking. However, it is recognized that, regardless of which method of producing a conjugate of the invention is selected, a determination may be needed to confirm that the peptide described herein maintains its antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity and that the moiety maintains its relevant function. Methods for determining the activity of the conjugates are well know to one of skill in the art.

[0127] An agar radial diffusion assay has been used as described herein to determine antimicrobial activities of proteins against microbial pathogens in vitro. One million CFU will be mixed into 10 ml (i.e., 1×10⁵ CFU/ml) of melted 1% agarose (in 10 mM NaHPO₄ and cooled to 42° C.) containing minimal nutrient and adjusted to either pH 5.5 or pH 7.2. The agar is solidified in culture dishes, and sample wells are formed. Peptides at various concentrations are dissolved in 10 μl of 0.01% acetic acid buffer (pH 5.5 or 7.2), loaded into individual wells, and incubated at 37° C. for three hours. The plate is then overlayed with 1% agarose containing nutrients and incubated (37° C., for at least 24 hours). Peptides purified by RP-HPLC lacking antimicrobial activity are tested in parallel as controls. Zones of inhibition are measured to quantify antimicrobial activity. This assay will not distinguish between microbicidal and microbiostatic actions, but is highly sensitive to peptides with one or both functions.

[0128] Minimum inhibitory (MIC) and microbiocidal concentration (MMC) assays can also be performed, and may include a microvolume assay which is used to quantitatively screen peptides for antimicrobial activities. In this assay, suspensions of bacteria or fungi in appropriate media are placed in 100-200 μl final volumes in microtiter plates. Poly-L-lysine coated or otherwise positively charged plates are used for these assays, since cationic peptides may bind to anionic surfaces. Purified peptides are then serially diluted, descending from 100 μg/ml. Organisms are inoculated into wells to a concentration of 1×10⁵ CFU/ml, and plates incubated (37° C., for at least 24 hours). Well turbidities are then assessed visually and by spectrophotometry to quantify growth inhibition versus wells containing no peptide. MMCs are then determined by quantitative culture of MIC wells exhibiting no visible growth.

[0129] Microbicidal kinetics of purified peptides are assessed by resuspending the peptides in 0.01% acetic acid buffer (pH 5.5 or 7.2), and organisms are resuspended to a concentration of 1×10⁵ CFU/ml in 50-250 μl of sterile buffer containing peptide concentrations from 0 to 40 μg/ml. Controls contain buffer alone or non-antimicrobial proteins and organism as above. Mixtures are incubated at 37° C. for up to 48 hours, after which aliquots are quantitatively cultured and incubated for 24 to 48 hours. Killing is expressed as decrease in logarithm₁₀ surviving CFU/ml. The limit of sensitivity in microbicidal assays is considered to be a 1 log reduction in viable cells.

[0130] Flow cytometry can also be used to examine kinetics and mechanisms of the action of the peptides on bacterial membrane integrity and energetics. Peptides which differ in activity or specificity for their ability to depolarize and/or permeabilize microbial membranes can also be compared by analysis of membrane depolarization, and permeabilization. DiOC₅ is a charged lipophilic dye which partitions into the cytoplasm, and is dependent on intact transmembrane potential (ΔΨ) for intracellular retention. Organisms prepared as above are labeled in darkness for 30 minutes at about 20° C. in PBS containing 0.05 μM DiOC₅ Organisms are resuspended to a concentration of 5×10⁸ CFU/ml in K.sup.+ MEM containing an individual peptide, and incubated at 37° C. For flow cytometry, organisms are washed, sonicated, counted, and resuspended in K.sup.+ MEM buffer. Reductions in mean DiOC₅ fluorescence relative to controls are interpreted to represent loss of DiOC₅, indicating membrane depolarization. Positive control cells exposed to valinomycin, as well as control cells not exposed to any peptides, are analyzed for DiOC₅ fluorescence in parallel.

[0131] Propidium iodide is excluded from cells with normal membrane integrity, but enters cells permealized to molecules ≧2 nm in diameter, and can be stimulated to emit fluorescence at >620 nm. Organisms prepared as above are resuspended to a concentration of 5×10⁸ CFU/ml in K.sup.+ MEM containing a selected peptide, and incubated for pre-selected times (ranging from zero up to about 120 minutes) at 37° C. Cells are washed in fresh K.sup.+ MEM, sonicated, counted, and resuspended in K.sup.+ MEM buffer containing 20 μM propidium iodide. Control cells exposed to ethanol (positive control for permeabilization) are assessed for propidium iodide uptake in parallel. Increases in mean propidium iodide fluorescence relative to control cells are interpreted to indicate increases in permeability.

[0132] Erythrocyte permeabilizing and hemolytic activities of peptides exhibiting potent microbicidal activity are also studied as indicators of potential in vivo toxicity. Four-percent (vol/vol) of washed human erythrocytes (in PBS alone, or in PBS plus 10% heat-inactivated PNHS are incubated with selected peptides ranging in concentration up to 100 times greater than geometric mean MICs. After 24 hours of incubation at 37° C., erythrocyte permeabilization and hemolysis are determined spectrophotometrically. Permeabilization and hemolysis will be compared to buffers alone, and with a triton X-100 control (100% hemolysis).

[0133] Endothelial cell injury due to peptides can also be measured using a standard ⁵¹Cr release assay, described in Filler et al., J Infect Dis., 164:928-935 (1991); Filler, et al., Infect Immun. 62:1064-1069 (1994); Filler et al., Infect Immun. 63:976-983 (1995). Briefly, endothelial cells in 96 well tissue culture plates are incubated with Na⁵¹ CrO₄ overnight. The following day, the unincorporated isotope tracer is removed by rinsing, and peptides in 0.01% acetic acid buffer are added to the endothelial cells. Control wells are exposed to buffer alone. After a predetermined incubation period, the medium is aspirated and the amount of ⁵¹Cr released into the medium is measured by scintillation. This approach facilitates toxicity screening of multiple peptides simultaneously, and minimizes the amount of peptide necessary for assessment.

[0134] Each antimicrobial and toxicity assay described above is performed independently a minimum of two times, and means±standard error is calculated for each peptide under varying exposure conditions (concentration or pH) as compared with control samples. Statistical analyses of microbicidal data are performed using Student t test or Kruskall-Wallis rank sum analysis for non-parametric data, and corrected for multiple comparisons as appropriate.

[0135] The yield of moiety/peptide conjugate formed is determined using routine methods. For example, HPLC or capillary electrophoresis or other qualitative or quantitative method can be used (see, for example, Liu et al., J. Chromatogr. 735:357-366 (1996); Rose et al., J. Chromatogr. 425:419-412 (1988), each of which is incorporated herein by reference). In particular, the skilled artisan will recognize that the choice of a method for determining yield of a conjugation reaction depends, in part, on the physical and chemical characteristics of the specific moiety and peptide. Following conjugation, the reaction products can be desalted to remove any free peptide and free drug.

[0136] Embodiments of the invention provide a method of inducing programmed cell death in a cell, including contacting the cell with an isolated peptide, protide or conjugate described herein. In one aspect, the cell is a microorganism, or in some aspects a pathogenic microorganism. In some aspects, the pathogenic microorganism is Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Bacillus subtilis, Acinotobacter baumannii, Acinotobacter calcoaceticus, Acinotobacter haemolyticus, Pseudomonas aeruginosa, Candida albicans. or any of the pathogenic microorganisms described herein. In another aspect, the cell is a tumor cell, including both a malignant and non-malignant tumor cell, or in a preferred aspect a malignant cell. In yet another aspect, the cell is an immune regulatory cell or an immune effector cell.

[0137] A peptide, protide or conjugate of the invention useful for practicing the methods of the invention can be formulated and administered by those skilled in the art in a manner and in an amount appropriate for the pathological condition to be treated, for example, an infection, neoplastic disorder, inflammation; the rate or amount of inflammation; the weight, gender, age and health of the individual; the biochemical nature, bioactivity, bioavailability and side effects of the particular compound; and in a manner compatible with concurrent treatment regimens. An appropriate amount and formulation for decreasing the severity of a pathological condition in humans can be extrapolated from credible animal models known in the art of the particular disorder. It is understood, that the dosage of a therapeutic substance has to be adjusted based on the binding affinity of the substance, such that a lower dose of a substance exhibiting significantly higher binding affinity can be administered compared to the dosage necessary for a substance with lower binding affinity. For a peptide, protide or conjugate described herein several factors can be taken into account when determining the proper dosage. For example, for a protide, the nature of the protide effectors and their bioactivity upon activation, the anticipated concentration of activator and the responsiveness of the activator site to presence of the activator, may be taken into account.

[0138] The total amount of peptide, protide or conjugate can be administered as a single dose or by infusion over a relatively short period of time, or can be administered in multiple doses administered over a more prolonged period of time. Such considerations will depend on a variety of case-specific factors such as, for example, whether the disease category is characterized by acute episodes or gradual or chronic deterioration. For an individual affected with an acute infection or inflammatory response, for example, as associated with a bacterial infection, the substance can be administered as a single dose or by infusion of several large doses in a relatively short period of time. For an individual affected with chronic deterioration, for example, as associated with a neuroinflammatory disorder, the substance can be administered in a slow-release matrice, which can be implanted for systemic delivery or at the site of the target tissue, which means an area proximal to the desired context. Contemplated matrices useful for controlled release of therapeutic compounds are well known in the art, and include materials such as DepoFoam®, biopolymers, micropumps, and the like.

[0139] The peptides, protides and conjugates administered in the methods of the invention can be administered to the individual by any number of routes known in the art including, for example, systemically, such as intravenously or intraarterially. A therapeutic peptide, protide or conjugate can be provided in the form of isolated and substantially purified polypeptides in pharmaceutically acceptable formulations using formulation methods known to those of ordinary skill in the art. These formulations can be administered by standard routes, including for example, topical, transdermal, intraperitoneal, intracranial, intracerebroventricular, intracerebral, intravaginal, intrauterine, oral, rectal or parenteral such as intravenous, intraspinal, intrathecal, subcutaneous or intramuscular routes. Intrathecal administration of a therapeutic peptide, protide or conjugate into the intradural or subarachnoid space can be an appropriate route for decreasing the severity of a neuroinflammatory condition. Intravenous administration of a therapeutic substance containing a peptide, protide or conjugate also is a preferred route for practicing the invention. In addition, a therapeutic substance administered in the methods of the invention can be incorporated into biodegradable polymers allowing for sustained release of the substance useful for prophylactic and reconstitutive applications described above. Biodegradable polymers and their use are described, for example, in Brem et al., J. Neurosurg. 74:441 446 (1991), which is incorporated herein by reference.

[0140] The methods for treating a particular pathological condition additionally can be practiced in conjunction with other therapies. For example, for treating cancer, the methods of the invention can be practiced prior to, during, or subsequent to conventional cancer treatments such as surgery, chemotherapy, including administration of cytokines and growth factors, radiation or other methods known in the art. Similarly, for treating pathological conditions which include infectious disease, the methods of the invention can be practiced prior to, during, or subsequent to conventional treatments, such as antibiotic administration, against infectious agents or other methods known in the art. Treatment of pathological conditions of autoimmune disorders also can be accomplished by combining the methods of the invention for inducing an immune response with conventional treatments for the particular autoimmune diseases. Conventional treatments include, for example, chemotherapy, steroid therapy, insulin and other growth factor and cytokine therapy, passive immunity and inhibitors of T cell receptor binding. The peptides, protides and conjugates of the invention can be administered in conjunction with these or other methods known in the art and at various times prior, during or subsequent to initiation of conventional treatments. For a description of treatments for pathological conditions characterized by aberrant cell growth see, for example, The Merck Manual, Sixteenth Ed, (Berkow, R., Editor) Rahway, N.J., 1992.

[0141] As described above, administration of a peptide, protide or conjugate can be, for example, simultaneous with or delivered in alternative administrations with the conventional therapy, including multiple administrations. Simultaneous administration can be, for example, together in the same formulation or in different formulations delivered at about the same time or immediately in sequence. Alternating administrations can be, for example, delivering a peptide, protide or conjugate formulation and a conventional therapeutic treatment in temporally separate administrations. Temporally separate administrations of a peptide, protide or conjugate and conventional therapy can use different modes of delivery and routes.

[0142] A therapeutic peptide, protide or conjugate containing substance administered in the methods of the invention also can be administered as a solution or suspension together with a pharmaceutically acceptable medium. Such a pharmaceutically acceptable medium can include, for example, sterile aqueous solvents such as sodium phosphate buffer, phosphate buffered saline, normal saline or Ringer's solution or other physiologically buffered saline, or other solvent or vehicle such as a glycol, glycerol, an oil such as olive oil or an injectable organic ester. A pharmaceutically acceptable medium can additionally contain physiologically acceptable compounds that act, for example, stabilize the neutralizing agent, increase its solubility, or increase its absorption. Such physiologically acceptable compounds include, for example, carbohydrates such as glucose, sucrose or dextrans; antioxidants such as ascorbic acid or glutathione; receptor mediated permeabilizers, which can be used to increase permeability of the blood-brain barrier; chelating agents such as EDTA, which disrupts microbial membranes; divalent metal ions such as calcium or magnesium; low molecular weight proteins; lipids or liposomes; or other stabilizers or excipients. Those skilled in the art understand that the choice of a pharmaceutically acceptable carrier depends on the route of administration of the compound containing the protides and on its particular physical and chemical characteristics.

[0143] Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions such as the pharmaceutically acceptable mediums described above. The solutions can additionally contain, for example, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient. Other formulations include, for example, aqueous and non-aqueous sterile suspensions which can include suspending agents and thickening agents. The formulations can be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and can be stored in a lyophilized condition requiring, for example, the addition of the sterile liquid carrier, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the kind previously described.

[0144] For applications that require the peptide, protide, or conjugate containing compounds to cross the blood-brain barrier, formulations that increase the lipophilicity of the compound are particularly desirable. For example, the neutralizing agent can be incorporated into liposomes (Gregoriadis, Liposome Technology, Vols. I to III, 2nd ed. (CRC Press, Boca Raton Fla. (1993)). Liposomes, which consist of phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0145] A therapeutic peptide, protide or conjugate containing substance administered in the methods of the invention can also be prepared as nanoparticles. Adsorbing peptide compounds onto the surface of nanoparticles has proven effective in delivering peptide drugs to the brain (see Kreuter et al., Brain Research 674:171 174 (1995)). Exemplary nanoparticles are colloidal polymer particles of poly-butylcyanoacrylate with a therapeutic protide-containing substance to be administered in the methods of the invention adsorbed onto the surface and then coated with polysorbate 80.

[0146] Image-guided ultrasound delivery of a therapeutic peptide, protide or conjugate containing substance administered in the methods of the invention through the blood-brain barrier to selected locations in the brain can be utilized as described in U.S. Pat. No. 5,752,515. Briefly, to deliver a therapeutic substance past the blood-brain barrier a selected location in the brain is targeted and ultrasound used to induce a change detectable by imaging in the central nervous system (CNS) tissues and/or fluids at that location. At least a portion of the brain in the vicinity of the selected location is imaged, for example, via magnetic resonance imaging (MRI), to confirm the location of the change. An therapeutic substance administered in the methods of the invention into the patient's bloodstream can be delivered to the confirmed location by applying ultrasound to effect opening of the blood-brain barrier at that location and, thereby, to induce uptake of the substance.

[0147] In addition, polypeptides called receptor mediated permeabilizers (RMP) can be used to increase the permeability of the blood-brain barrier to molecules such as therapeutic, prophylactic or diagnostic substances as described in U.S. Pat. Nos. 5,268,164; 5,506,206; and 5,686,416. These receptor mediated permeabilizers can be intravenously co-administered to a host with molecules whose desired destination is the cerebrospinal fluid compartment of the brain, for example, in the treatment of a neuroinflammatory condition. The permeabilizer polypeptides or conformational analogues thereof allow therapeutic substances to penetrate the blood-brain barrier and arrive at their target destination which can be selected based on its proximity to the desired activation context. Such polypeptides can be designed as part of strategic invention protides.

[0148] In current treatment regimes for most diseases, more than one compound is often administered to an individual for management of the same or different aspects of the disease. Similarly, in the methods of the invention for treating neoplastic condition, microbial infection, a condition associated with decreased cell death or inflammatory condition, a therapeutic peptide, protide or conjugate containing substance can advantageously be formulated with a second therapeutic compound such as an anti-inflammatory compound, antimicrobial compound, chemotherapeutic compound, immunosuppressive compound or any other compound that manages the same or different aspects of the particular disease. As an example, for treatment of an infectious disease a therapeutic substance can advantageously be formulated with a second therapeutic compound such as an antibiotic. Contemplated methods of treating a pathological condition by administering to a subject a therapeutically effective amount of a peptide, protide or conjugate therefore include administering a therapeutic substance useful in the methods of the invention alone, in combination with, or in sequence with, such other compounds. Alternatively, combination therapies can consist of fusion proteins, where a therapeutic substance useful for treating a particular pathological condition is linked to a heterologous protein, such as an invention protide.

[0149] It is understood that modifications which do not substantially affect the activity of the various embodiments of this invention are also provided within the definition of the invention provided herein. Accordingly, the following examples are intended to illustrate but not limit the present invention.

Example I

Antimicrobial, Anti-Cancer, and Other Therapeutic Applications of Peptides Designed from Programmed Cell Death Effector Proteins

[0150] Candidate proteins representing a variety of nuclear- or mitochondrial-encoded programmed cell death effector and/or modulatory molecules from eukaryotic sources were identified for comparison. When candidate proteins were compared, similarities in sequences and helical domains that have amphipathic and cationic characteristics were identified. Analogues were also identified where possible from prokaryotic sources.

[0151] Template proteins were initially chosen based on the following: 1) mammalian or other eukaryotic cell nuclear-encoded peptides with structure-activity relationships to antimicrobial peptide sequences that afford interaction/control/inhibition of mitochondria or chloroplast symbionts, or host cell apoptosis (examples include Bax, Bcl-W, dynamin, mitofusin, nucleolin, and other programmed cell death effector proteins); and 2) structural and/or functional homologues of such proteins in prokaryotes (examples include CidA/B, LrgA/B, and other holin-like or programmed cell death effector proteins). The identification of candidate peptide sequences was achieved through an iterative process, which included a search of protein databases for additional candidate proteins, multiseqeunce alignments of the identified candidates, followed by an integration and analysis of candidate sequences.

[0152] Upon identification of an initial group of template proteins having the above-identified characteristics, additional candidate proteins were identified using primary sequence similarity searches of protein databases using the basic local alignment search tool (BLAST) available from the National Center for Biotechnology Information. The searches utilized the amino acid sequences of candidate template proteins having the desired characteristics as query sequences. FIG. 3 shows exemplary amino acid sequence used as the query sequences.

[0153] Once all additional candidate template proteins were identified, a compiled set of query sequences were submitted for analysis to the modeling server ClustalW (Larkin et al., Bioinformatics 23(21): 2947-2948 (2007)) available online from EMBL-EBI. Utilizing the modeling server, multisequence alignments were performed to identify regions of sequence homology within the candidate sequences or with known host defense or antimicrobial peptide sequences. For example, candidate proteins were ranked based on their alignment score (FIG. 4) and/or analyzed for conserved residues through multisequence alignments (FIG. 5). Additionally, phylogenetic and cladogenetic analyses were conducted between the candidate proteins, followed by multiseqeunce alignments of identified putative helical domains (FIGS. 6-14).

[0154] The results from the above processes were integrated to analyze and prioritize candidate sequences. The criteria for their prioritization included: 1) conservation of sequence homology or motif(s); 2) homology to known antimicrobial or anti-cancer peptides; 3) similarity to known or recognized antimicrobial peptide structure-activity relationships (SAR; including presence, periodicity, and distribution of cationic, hydrophobic, and aromatic residues); and 4) visual inspection. Candidate sequences that were identified included mitochondria-targeting proteins, such as Bcl-2, Bcl-W, Bax, and Mitofusin; NK/Tcyto cell effectors, such as Granulysin, Granzyme H, Perforin-1 and Azurocidin (CAP37); apoptosis/cell signaling proteins, such as Fas ligand, Caspase 7 and Dynamin 1; and other related proteins, such as Serpin B9 (CytoPro3), Fractalkine (CX3CL1), CXCL3 and Atrophin 1.

[0155] Following the above interactive primary structure analysis, a secondary (2°) structure analysis was conducted. This analysis included visualization, qualitative, and/or quantitative analyses of candidate sequences identified above. This analysis, including 3D visualization of target sequences, conformation 3D homology, qualitative 3D analysis of target sequences, quantitative 3D analysis of target sequences and comparative 3D refinement of target sequences.

[0156] 3D visualization of target sequences was achieved using Cn3D software available through PubMed. Exemplary 3D visualization of identified candidate peptides within the native total protein are shown in FIG. 15-21. These results identified specific sequences as novel targets for further analysis/design. The conformational 3D homology of target sequences to known antimicrobial or anti-cancer template polypeptides was assessed using the threading and 3D homology fold recognition server Protein Homology/analogy Recognition Engine (PHYRE) "Protein structure prediction on the web: a case study using the Phyre server" (Kelley and Sternberg Nature Protocols 4:363-371 (2009). Statistical e values were used to guide prioritization of molecules for further analysis.

[0157] Priority target sequences identified above were visualized for qualitative analysis of distribution of 3D physicochemical attributes using the public domain UCSF software package Chimera. Next, priority target sequences were quantitatively evaluated for structural homology and/or structure-activity relationships to known antimicrobial and/or anti-cancer peptides/proteins using a combinatorial extension method of Shindyalov and Bourne, Protein Engineering 11:739-747 (1998). The results from these analyses provided quantitative alignment of compositional elements, including charged and hydrophobic residues (FIGS. 22-29). The results included root mean square deviation (RMSD) scores as quantitative data that allowed further prioritization of target sequences. The priority target sequences emanating from the process described above were then used as novel templates for 3D analyses using VAST and/or 3Dpssm software to identify homologous sequences and discover other novel target sequences.

[0158] As a final step, a computation simulation of the antimicrobial activity of the selected sequences was conducted. Selected target sequences emerging from the above process were subjected to a computational assessment tool which integrates multiple physical and biochemical attributes of polypeptide sequences to generate a predicted minimal inhibitory concentration (MIC) based on the inverse of the target sequence calculated hydrophobic moment (1/MH; see U.S. Pat. No. 6,743,769).

[0159] Based on the above process, specific peptide sequences were identified, which have the desired primary and secondary structure within the candidate proteins (Tables 1-20). These identified peptides are predicted to have antimicrobial and anti-cancer activity. Exemplifying predictive accuracy, candidate peptides have been synthesized and antimicrobial efficacy has been demonstrated in vitro against a panel of Gram-positive and Gram-negative bacteria and fungi (see Example II). Furthermore, specific amino acid substitutions were identified for several peptide sequences (Tables 1-19). The identified peptide residues were prioritized/ranked utilizing to above process (Table 21).

TABLE-US-00001 TABLE 1 Engineered peptides based on Bax protein (pro-apoptotic protein/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 1. ¹⁰4nfnwgrvvalfyfasklvlkalctkv¹²⁹ 2. .sup.168twqtvtifvagvltasltiwkk¹⁹⁰ Synthetic Peptides 1. H₂N-nfnwgrvvalfyfasklvlkalctkv-COOH 1. H₂N-nfnwgrvvalfyfasklvlkalctkv-CONH₂ 3. H₂N-nfnwgrvvalfyfasklvlkalxtkv-COOH 3. H₂N-nfnwgrvvalfyfasklvlkalxtkv-CONH₂ 4. H₂N-nfnwgrvvalfyfasklvlkalbtjv-COOH 4. H₂N-nfnwgrvvalfyfasklvlkalbtjv-CONH₂ 5. H₂N-wgrvvalfyfasklvlkalctkv-COOH 5. H₂N-wgrvvalfyfasklvlkalctkv-CONH₂ 6. H₂N-wgrvvalfyfasklvlkalxtkv-COOH 6. H₂N-wgrvvalfyfasklvlkalxtkv-CONH₂ 7. H₂N-rvvalfyfasklvlkalctkv-COOH 7. H₂N-rvvalfyfasklvlkalctkv-CONH₂ 8. H₂N-rvvalfyfasklvlkalxtkv-COOH 8. H₂N-rvvalfyfasklvlkalxtkv-CONH₂ 9. H₂N-alfyfasklvlkalctkv-COOH 9. H₂N-alfyfasklvlkalctkv-CONH₂ 10. H₂N-alfyfasklvlkalxtkv-COOH 10. H₂N-alfyfasklvlkalxtkv-CONH₂ 2. H₂N-twqtvtifvagvltasltiwkk-COOH 2. H₂N-twqtvtifvagvltasltiwkk-CONH₂ 11. H₂N-twqtvtifvabvltasltiwkk-COOH 11. H₂N-twqtvtifvabvltasltiwkk-CONH₂ 12. H₂N-tvtifvagvltasltiwkk-COOH 12. H₂N-tvtifvagvltasltiwkk-CONH₂ 13. H₂N-tvtifvabvltasltiwkk-COOH 13. H₂N-tvtifvabvltasltiwkk-CONH₂ Underlined residues indicate substitutions; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue; b = k, r, n, q, other natural/non-natural basic residue; j = cysteine or other natural/non-natural thiol residue.

TABLE-US-00002 TABLE 2 Engineered peptides based on Bcl-W protein (pro-apoptotic protein/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 14. ¹⁰tralvadfvgyklrqkgyv²⁸ 15. ⁵⁵trfrrtfsdlaaqlhvt⁷¹ 16. .sup.159arrlregnwasvrtvltgavalgalvtvgaffask¹⁹³ Synthetic Peptides 14. H₂N-tralvadfvgyklrqkgyv-COOH 14. H₂N-tralvadfvgyklrqkgyv-CONH₂ 17. H₂N-tralvabfvgyklrqkgyv-COOH 17. H₂N-tralvabfvgyklrqkgyv-CONH₂ 15. H₂N-trfrrtfsdlaaqlhvt-COOH 15. H₂N-trfrrtfsdlaaqlhvt-CONH₂ 18. H₂N-trfrbtfsdlaaqlhvt-COOH 18. H₂N-trfrbtfsdlaaqlhvt-CONH₂ 19. H₂N-trfrrtfsblaaqlhvt-COOH 19. H₂N-trfrrtfsblaaqlhvt-CONH₂ 20. H₂N-rrlregnwasvrtvltgavalgalvtvgaffask-COOH 20. H₂N-rrlregnwasvrtvltgavalgalvtvgaffask-CONH₂ 21. H₂N-rrlrbgnwasvrtvltgavalgalvtvgaffask-COOH 21. H₂N-rrlrbgnwasvrtvltgavalgalvtvgaffask-CONH₂ 22. H₂N-rrlregnwasvrtvltbavalgalvtvgaffask-COOH 22. H₂N-rrlregnwasvrtvltbavalgalvtvgaffask-CONH₂ 23. H₂N-rrlregnwasvrtvltgavalbalvtvgaffask-COOH 23. H₂N-rrlregnwasvrtvltgavalbalvtvgaffask-CONH₂ 24. H₂N-rrlregnwasvrtvltgavalgalvtvbaffask-COOH 24. H₂N-rrlregnwasvrtvltgavalgalvtvbaffask-CONH₂ 25. H₂N-rrlrbgnwasvrtvltbavalbalvtvbaffask-COOH 25. H₂N-rrlrbgnwasvrtvltbavalbalvtvbaffask-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue.

TABLE-US-00003 TABLE 3 Engineered peptides based on Bcl-xβ protein (apoptotic protein/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 26. ¹²flsyklsqkgyswsqfs²⁸ 27. ⁹⁹lryrrafsdltsqlhitpgtayqsf¹²² 28. .sup.187gwvrtkplvcpfslasgqrs¹⁹⁶ 29. .sup.203gqrsptalllylfllcwvivg²²³ Synthetic Peptides 26. H₂N-flsyklsqkgyswsqfs-COOH 26. H₂N-flsyklsqkgyswsqfs-CONH₂ 27. H₂N-lryrrafsdltsqlhitpgtayqsf-COOH 27. H₂N-lryrrafsdltsqlhitpgtayqsf-CONH₂ 30. H₂N-lryrrafsbltsqlhitpgtayqsf-COOH 30. H₂N-lryrrafsbltsqlhitpgtayqsf-CONH₂ 28. H₂N-gwvrtkplvcpfslasgqrs-COOH 28. H₂N-gwvrtkplvcpfslasgqrs-CONH₂ 31. H₂N-gwvrtkplvxpfslasgqrs-COOH 31. H₂N-gwvrtkplvxpfslasgqrs-CONH₂ 32. H₂N-gwvrtkplvxpfslasbqrs-COOH 32. H₂N-gwvrtkplvxpfslasbqrs-CONH₂ 29. H₂N-gqrsptalllylfllcwvivg-COOH 29. H₂N-gqrsptalllylfllcwvivg-CONH₂ 33. H₂N-gqrsptalxlylfllcwvivg-COOH 33. H₂N-gqrsptalxlylfllcwvivg-CONH₂ 34. H₂N-gqrsptalllylfllxwvivg-COOH 34. H₂N-gqrsptalllylfllxwvivg-CONH₂ 35. H₂N-gqrsptalllylfllcwvivb-COOH 35. H₂N-gqrsptalllylfllcwvivb-CONH₂ 36. H₂N-gqrsptalxlylfllxwvivb-COOH 36. H₂N-gqrsptalxlylfllxwvivb-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00004 TABLE 4 Engineered peptides based on Bak protein (apoptotic protein/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 37. ¹²⁷rvvallgfgyrlalhvyq¹⁴⁴ 38. .sup.187ilnvlvvlgvvllgqfvvrrffks²¹¹ Synthetic Peptides 37. H₂N-rvvallgfgyrlalhvyq-COOH 37. H₂N-rvvallgfgyrlalhvyq-CONH₂ 39. H₂N-rvvallbfgyrlalhvyq-COOH 39 H₂N-rvvallbfgyrlalhvyq-CONH₂ 40. H₂N-rvvallgfbyrlalhvyq-COOH 40. H₂N-rvvallgfbyrlalhvyq-CONH₂ 41. H₂N-rvvallbfbyrlalhvyq-COOH 41. H₂N-rvvallbfbyrlalhvyq-CONH₂ 42. H₂N-rvvalygfgyrlalhvyq-COOH 42. H₂N-rvvalygfgyrlalhvyq-CONH₂ 43. H₂N-rvvalwgfgyrlalhvyq-COOH 43. H₂N-rvvalwgfgyrlalhvyq-CONH₂ 44. H₂N-rvvalybfbyrlalhvyq-COOH 44. H₂N-rvvalybfbyrlalhvyq-CONH₂ 45. H₂N-rvvalwbfbyrlalhvyq-COOH 45. H₂N-rvvalwbfbyrlalhvyq-CONH₂ 38. H₂N-ilnvlvvlgvvllgqfvvrrffks-COOH 38. H₂N-ilnvlvvlgvvllgqfvvrrffks-CONH₂ 46. H₂N-ilnvlvxlgvvllgqfvvrrffks-COOH 46. H₂N-ilnvlvxlgvvllgqfvvrrffks-CONH₂ 47. H₂N-ilnvlvblvlgqfvrfks-COOH 47. H₂N-ilnvlvblvlgqfvrfks-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00005 TABLE 5 Engineered peptides based on Bcl-2 protein (apoptotic regulator/human). SEQ ID NO: Amino Acid Sequence Native Sequence Domain 48. ¹¹nreivmkyihyklsqrgy²⁸ Synthetic Peptides 48. H₂N-nreivmkyihyklsqrgy-COOH 48. H₂N-nreivmkyihyklsqrgy-CONH₂ 49. H₂N-nrbivmkyihyklsqrgy-COOH 49. H₂N-nrbivmkyihyklsqrgy-CONH₂ 50. H₂N-nreivxkyihyklsqrgy-COOH 50. H₂N-nreivxkyihyklsqrgy-CONH₂ 51. H₂N-nreivmkyibyklsqrgy-COOH 51. H₂N-nreivmkyibyklsqrgy-CONH₂ 52. H₂N-nrbivxkyibyklsqrgy-COOH 52. H₂N-nrbivxkyibyklsqrgy-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor residue.

TABLE-US-00006 TABLE 6 Engineered peptides based on Bcl-2 isoform 1 protein (apoptotic regulator/human). SEQ ID NO: Amino Acid Sequence Native Sequence Domain 53. ⁵⁴lalrqagddfsrryrg⁶⁹ Synthetic Peptides 53. H₂N-lalrqagddfsrryrg-COOH 53. H₂N-lalrqagddfsrryrg-CONH₂ 54. H₂N-lalrqagbdfsrryrg-COOH 54. H₂N-lalrqagbdfsrryrg-CONH₂ 55. H₂N-lalrqagdbfsrryrg-COOH 55. H₂N-lalrqagdbfsrryrg-CONH₂ 56. H₂N-lalrqagbxfsrryrg-COOH 56. H₂N-lalrqagbxfsrryrg-CONH₂ 57. H₂N-lalrqaobxfsrryrg-COOH 57. H₂N-lalrqaobxfsrryrg-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue; o = anthrylalanine or other non-natural amino acid.

TABLE-US-00007 TABLE 7 Engineered peptides based on Mfn-1 protein (mitofusin-1; human mitochondrial regulator). SEQ ID NO: Amino Acid Sequence Native Sequence Domain 58. ⁶⁹9keidqlekiqnnskllrnkavqlenelenftkqfl⁷³⁴ Synthetic Peptides 58. H₂N-keidqlekiqnnskllrnkavqlenelenftkqfl-COOH 58. H₂N-keidqlekiqnnskllrnkavqlenelenftkqfl-CONH₂ 59. H₂N-keibqlekiqnnskllrnkavqlenelenftkqfl-COOH 59 H₂N-keibqlekiqnnskllrnkavqlenelenftkqfl-CONH₂ 60. H₂N-keibqlbkiqnnskllrnkavqlbnelenftkqfl-COOH 60. H₂N-keibqlbkiqnnskllrnkavqlbnelenftkqfl-CONH₂ 61. H₂N-keibqlbkiqnnskllrnkavqlbnelenftkqfl-COOH 61. H₂N-keibqlbkiqnnskllrnkavqlbnelenftkqfl-CONH₂ 62. H₂N-keibqlbkiqnnskllrnkavqlbnblenftkqfl-COOH 62. H₂N-keibqlbkiqnnskllrnkavqlbnblenftkqfl-CONH₂ 63. H₂N-keibqlbkiqnnskllrnkavqlbnblbnftkqfl-COOH 63. H₂N-keibqlbkiqnnskllrnkavqlbnblbnftkqfl-CONH₂ 64. H₂N-kiqnnskllrnkavql-COOH 64. H₂N-kiqnnskllrnkavql-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue.

TABLE-US-00008 TABLE 8 Engineered peptides based on Mfn-2 protein (mitofusin-1; human mitochondrial regulator). SEQ ID NO: Amino Acid Sequence Native Sequence Domain 65. ⁷¹8nkkievldslqskakllrnkagwldselnmfthqylqpsr⁷⁵⁷ Synthetic Peptides 65. H₂N-nkkievldslqskakllrnkagwldselnmfthqylqpsr-COOH 65. H₂N-nkkievldslqskakllrnkagwldselnmfthqylqpsr-CONH₂ 66. H₂N-nkkibvldslqskakllrnkagwldselnmfthqylqpsr-COOH 66. H₂N-nkkibvldslqskakllrnkagwldselnmfthqylqpsr-CONH₂ 67. H₂N-nkkievlbslqskakllrnkagwldselnmfthqylqpsr-COOH 67. H₂N-nkkievlbslqskakllrnkagwldselnmfthqylqpsr-CONH₂ 68. H₂N-nkkievldslqskakllrnkagwlbselnmfthqylqpsr-COOH 68. H₂N-nkkievldslqskakllrnkagwlbselnmfthqylqpsr-CONH₂ 69. H₂N-nkkievldslqskakllrnkagwldsblnmfthqylqpsr-COOH 69. H₂N-nkkievldslqskakllrnkagwldsblnmfthqylqpsr-CONH₂ 70. H₂N-nkkievldslqskakllrnkagwldselnxfthqylqpsr-COOH 70. H₂N-nkkievldslqskakllrnkagwldselnxfthqylqpsr-CONH₂ 71. H₂N-nkkibvlbslqskakllrnkagwlbsblnxfthqylqpsr-COOH 71. H₂N-nkkibvlbslqskakllrnkagwlbsblnxfthqylqpsr-CONH₂ 72. H₂N-kkievldslqskakllrnkagwl-COOH 72. H₂N-kkievldslqskakllrnkagwl-CONH₂ 73. H₂N-kkibvldslqskakllrnkagwl-COOH 73. H₂N-kkibvldslqskakllrnkagwl-CONH₂ 74. H₂N-kkievlbslqskakllrnkagwl-COOH 74. H₂N-kkievlbslqskakllrnkagwl-CONH₂ 75. H₂N-kkibvlbslqskakllrnkagwl-COOH 75. H₂N-kkibvlbslqskakllrnkagwl-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00009 TABLE 9 Engineered peptides based on Dnm-1 protein (dynamin-1; human mitochondrial regulator). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 76. ²²³kllplrrgyigvvnrsqk²⁴⁰ 77. .sup.256rkfflshpsyrhla²⁶⁹ 78. .sup.294glrnklqsqllsiek³09 79. ³²3arktkallqmvqqfavdf³40 80. .sup.411atvkkqvqklk⁵⁰1 81. ⁵¹9virkgwltinnigimkggsk⁵39 82. ⁵69nlklrdvekgfmsskhifalfnteqrnvyk⁵⁹⁸ 83. .sup.617kasflragvypervgdk⁶³³ Synthetic Peptides 76. H₂N-kllplrrgyigvvnrsqk-COOH 76. H₂N-kllplrrgyigvvnrsqk-CONH₂ 77. H₂N-rkfflshpsyrhla-COOH 77. H₂N-rkfflshpsyrhla-CONH₂ 78. H₂N-glrnklqsqllsiek-COOH 78. H₂N-glrnklqsqllsiek-CONH₂ 84. H₂N-glrnklqsqllsibk-COOH 84. H₂N-glrnklqsqllsibk-CONH₂ 79. H₂N-arktkallqmvqqfavdf-COOH 79. H₂N-arktkallqmvqqfavdf-CONH₂ 85. H₂N-arktkallqmvqqfavbf-COOH 85. H₂N-arktkallqmvqqfavbf-CONH₂ 80. H₂N-atvkkqvqklk-COOH 80. H₂N-atvkkqvqklk-CONH₂ 81. H₂N-virkgwltinnigimkggsk-COOH 81. H₂N-virkgwltinnigimkggsk-CONH₂ 82. H₂N-nlklrdvekgfmsskhifalfnteqrnvyk-COOH 82. H₂N-nlklrdvekgfmsskhifalfnteqrnvyk-CONH₂ 86. H₂N-nlklrbvekgfmsskhifalfnteqrnvyk-COOH 86. H₂N-nlklrbvekgfmsskhifalfnteqrnvyk-CONH₂ 87. H₂N-nlklrdvbkgfmsskhifalfnteqrnvyk-COOH 87. H₂N-nlklrdvbkgfmsskhifalfnteqrnvyk-CONH₂ 88. H₂N-nlklrdvekgfmsskhifalfntbqrnvyk-COOH 88. H₂N-nlklrdvekgfmsskhifalfntbqrnvyk-CONH₂ 89. H₂N-nlklrbvbkgfmsskhifalfnteqrnvyk-COOH 89. H₂N-nlklrbvbkgfmsskhifalfnteqrnvyk-CONH₂ 90. H₂N-nlklrbvbkgfmsskhifalfntbqrnvyk-COOH 90. H₂N-nlklrbvbkgfmsskhifalfntbqrnvyk-CONH₂ 83. H₂N-kasflragvypervgdk-COOH 83. H₂N-kasflragvypervgdk-CONH₂ 91. H₂N-kasflragvypbrvgdk-COOH 91. H₂N-kasflragvypbrvgdk-CONH₂ 92. H₂N-kasflragvypervgbk-COOH 92. H₂N-kasflragvypervgbk-CONH₂ 93. H₂N-kasflragvypbrvgbk-COOH 93. H₂N-kasflragvypbrvgbk-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue.

TABLE-US-00010 TABLE 10 Engineered peptides based on Dnm-2 protein (dynamin-2; human mitochondrial regulator). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 94. .sup.187klakevdpqglrtigvitkl.sup.206 95. .sup.295alrsklqsqllslek³09 96. ³²3trktkallqmvqqfgv³38 97. .sup.420aivkkqvvklk.sup.421 98. ⁴⁹9aqqrstqlnkkraipnqg⁵¹6 99. ⁵⁵9kekkymlpldnlkirdvekgfmsnkhvfaifnteqrnvyk⁵⁹⁸ 100. ⁵69nlkirdvekgfmsnkhvfaifnteqrnvyk⁵⁹⁸ 101. .sup.615swkasflragvypekdqa⁶³² Synthetic Peptides 94. H₂N-klakevdpqglrtigvitkl-COOH 94. H₂N-klakevdpqglrtigvitkl-CONH₂ 102. H₂N-klakbvdpqglrtigvitkl-COOH 102. H₂N-klakbvdpqglrtigvitkl-CONH₂ 103. H₂N-klakevbpqglrtigvitkl-COOH 103. H₂N-klakevbpqglrtigvitkl-CONH₂ 104. H₂N-klakbvbpqglrtigvitkl-COOH 104. H₂N-klakbvbpqglrtigvitkl-CONH₂ 95. H₂N-alrsklqsqllslek-COOH 95. H₂N-alrsklqsqllslek-CONH₂ 105. H₂N-alrsklqsqllslbk-COOH 105 H₂N-alrsklqsqllslbk-CONH₂ 96. H₂N-trktkallqmvqqfgv-COOH 96. H₂N-trktkallqmvqqfgv-CONH₂ 97. H₂N-aivkkqvvklk-COOH 97. H₂N-aivkkqvvklk-CONH₂ 106. H₂N-oivkkqvvklk-COOH 106. H₂N-oivkkqvvklk-CONH₂ 98. H₂N-aqqrstqlnkkraipnqg-COOH 98. H₂N-aqqrstqlnkkraipnqg-CONH₂ 99. H₂N-kekkymlpldnlkirdvekgfmsnkhvfaifnteqrnvyk-COOH 99. H₂N-kekkymlpldnlkirdvekgfmsnkhvfaifnteqrnvyk-CONH₂ 107. H₂N-kekkymlpldnlkir-COOH 107. H₂N-kekkymlpldnlkir-CONH₂ 108. H₂N-kxkkymlpldnlkir-COOH 108. H₂N-kxkkymlpldnlkir-CONH₂ 109. H₂N-kekkyxlpldnlkir-COOH 109. H₂N-kekkyxlpldnlkir-CONH₂ 110. H₂N-kekkymlplbnlkir-COOH 110. H₂N-kekkymlplbnlkir-CONH₂ 111. H₂N-kekkymlplxnlkir-COOH 111. H₂N-kekkymlplxnlkir-CONH₂ 112. H₂N-kxkkymlplbnlkir-COOH 112. H₂N-kxkkymlplbnlkir-CONH₂ 113. H₂N-kxkkyxlplbnlkir-COOH 113. H₂N-kxkkyxlplbnlkir-CONH₂ 114. H₂N-kgfmsnkhvfaifnteqrnvyk-COOH 114. H₂N-kgfmsnkhvfaifnteqrnvyk-CONH₂ 115. H₂N-kxkkymlpldnlkirdvekgfmsnkhvfaifnteqrnvyk-COOH 115. H₂N-kxkkymlpldnlkirdvekgfmsnkhvfaifnteqrnvyk-CONH₂ 116. H₂N-kekkyxlpldnlkirdvekgfmsnkhvfaifnteqrnvyk-COOH 116. H₂N-kekkyxlpldnlkirdvekgfmsnkhvfaifnteqrnvyk-CONH₂ 117. H₂N-kekkymlpldnlkirdvekgfxsnkhvfaifnteqrnvyk-COOH 117. H₂N-kekkymlpldnlkirdvekgfxsnkhvfaifnteqrnvyk-CONH₂ 100. H₂N-nlkirdvekgfmsnkhvfaifnteqrnvyk-COOH 100. H₂N-nlkirdvekgfmsnkhvfaifnteqrnvyk-CONH₂ 118. H₂N-nlkirbvekgfmsnkhvfaifnteqrnvyk-COOH 118. H₂N-nlkirbvekgfmsnkhvfaifnteqrnvyk-CONH₂ 119. H₂N-nlkirdvbkgfmsnkhvfaifnteqrnvyk-COOH 119. H₂N-nlkirdvbkgfmsnkhvfaifnteqrnvyk-CONH₂ 120. H₂N-nlkirbvbkgfmsnkhvfaifntbqrnvyk-COOH 120. H₂N-nlkirbvbkgfmsnkhvfaifntbqrnvyk-CONH₂ 101. H₂N-swkasflragvypekdqa-COOH 101. H₂N-swkasflragvypekdqa-CONH₂ 121. H₂N-swkasflragvypbkdqa-COOH 121. H₂N-swkasflragvypbkdqa-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; o = anthrylalanine or other non-natural amino acid; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00011 TABLE 11 Engineered peptides based on Ncl protein (nucleolin; human mitochondrial regulator). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 122. ⁵⁰qkkgkkaaatsakkvvvs⁶⁷ 123. ⁶⁹tkkvavatpakkaavt⁸⁴ 124. ¹²4kkgaaipakgakngknakk¹⁴² 125. .sup.216akgkkaakvvpvkaknva²³³ 126. ²⁷3vkeapgkrkkemakqkaa²⁹⁰ 127. ³62kaleltglkvfgneiklek³80 128. ³82kgkdskkerdartllaknlpykvtq⁴⁰⁶ 129. .sup.419irlvskdgkskgiayi⁴³⁴ 130. .sup.467kgqnqdyrggknstwsgesktlvlsnlsysat⁴⁹⁸ 131. ⁵⁰8katfikvpqnqngkskgyafi⁵²⁸ Synthetic Peptides 122. H₂N-qkkgkkaaatsakkvvvs-COOH 122. H₂N-qkkgkkaaatsakkvvvs-CONH₂ 132. H₂N-qkkgkkaxatsakkvyvs-COOH 132. H₂N-qkkgkkaxatsakkvyvs-CONH₂ 133. H₂N-qkkgkkaaatsakkvyvs-COOH 133. H₂N-qkkgkkaaatsakkvyvs-CONH₂ 134. H₂N-qkkgkkaaatsakkvwvs-COOH 134. H₂N-qkkgkkaaatsakkvwvs-CONH₂ 135. H₂N-qkkgkkaxatsakkvyvs-COOH 135. H₂N-qkkgkkaxatsakkvyvs-CONH₂ 136. H₂N-qkkgkkaxatsakkvwvs-COOH 136. H₂N-qkkgkkaxatsakkvwvs-CONH₂ 123. H₂N-tkkvavatpakkaavt-COOH 123. H₂N-tkkvavatpakkaavt-CONH₂ 124. H₂N-kkgaaipakgakngknakk-COOH 124. H₂N-kkgaaipakgakngknakk-CONH₂ 137. H₂N-kkgaxipakgakngknakk-COOH 137. H₂N-kkgaxipakgakngknakk-CONH₂ 125. H₂N-akgkkaakvvpvkaknva-COOH 125. H₂N-akgkkaakvvpvkaknva-CONH₂ 138. H₂N-akgkkaakvvxvkaknva-COOH 138. H₂N-akgkkaakvvxvkaknva-CONH₂ 126. H₂N-vkeapgkrkkemakqkaa-COOH 126. H₂N-vkeapgkrkkemakqkaa-CONH₂ 139. H₂N-vkbapgkrkkemakqkaa-COOH 139 H₂N-vkbapgkrkkemakqkaa-CONH₂ 140. H₂N-vkeapgkrkkbmakqkaa-COOH 140. H₂N-vkeapgkrkkbmakqkaa-CONH₂ 141. H₂N-vkeapgkrkkexakqkaa-COOH 141. H₂N-vkeapgkrkkexakqkaa-CONH₂ 142. H₂N-vkbapgkrkkbxakqkaa-COOH 142. H₂N-vkbapgkrkkbxakqkaa-CONH₂ 127. H₂N-kaleltglkvfgneiklek-COOH 127. H₂N-kaleltglkvfgneiklek-CONH₂ 143. H₂N-kalbltglkvfgneiklek-COOH 143. H₂N-kalbltglkvfgneiklek-CONH₂ 144. H₂N-kaleltglkvfgnbiklek-COOH 144. H₂N-kaleltglkvfgnbiklek-CONH₂ 145. H₂N-kaleltglkvfgneiklbk-COOH 145. H₂N-kaleltglkvfgneiklbk-CONH₂ 146. H₂N-kalbltglkvfgnbiklbk-COOH 146. H₂N-kalbltglkvfgnbiklbk-CONH₂ 128. H₂N-kgkdskkerdartllaknlpykvtq-COOH 128. H₂N-kgkdskkerdartllaknlpykvtq-CONH₂ 147. H₂N-kgkxskkerdartllaknlpykvtq-COOH 147. H₂N-kgkxskkerdartllaknlpykvtq-CONH₂ 148. H₂N-kgkdskkxrdartllaknlpykvtq-COOH 148. H₂N-kgkdskkxrdartllaknlpykvtq-CONH₂ 149. H₂N-kgkdskkerbartllaknlpykvtq-COOH 149. H₂N-kgkdskkerbartllaknlpykvtq-CONH₂ 150. H₂N-kgkxskkbrbartllaknlpykvtq-COOH 150. H₂N-kgkxskkbrbartllaknlpykvtq-CONH₂ 129. H₂N-irlvskdgkskgiayi-COOH 129. H₂N-irlvskdgkskgiayi-CONH₂ 151. H₂N-irlvskfgkskgiayi-COOH 151. H₂N-irlvskfgkskgiayi-CONH₂ 152. H₂N-irlvskygkskgiayi-COOH 152. H₂N-irlvskygkskgiayi-CONH₂ 153. H₂N-irlvskwgkskgiayi-COOH 153 H₂N-irlvskwgkskgiayi-CONH₂ 154. H₂N-irlvsklwgkskgiayi-COOH 154 H₂N-irlvsklwgkskgiayi-CONH₂ 155. H₂N-irlvskdgkskg-COOH 155. H₂N-irlvskdgkskg-CONH₂ 156. H₂N-irlvskfgkskgi-COOH 156. H₂N-irlvskfgkskg-CONH₂ 157. H₂N-irlvskygkskg-COOH 157. H₂N-irlvskygkskg-CONH₂ 158. H₂N-irlvskwgkskg-COOH 158. H₂N-irlvskwgkskg-CONH₂ 159. H₂N-irlvsklwgkskg-COOH 159. H₂N-irlvsklwgkskgi-CONH₂ 130. H₂N-kgqnqdyrggknstwsgesktlvlsnlsysat-COOH 130. H₂N-kgqnqdyrggknstwsgesktlvlsnlsysat-CONH₂ 160. H₂N-kgqnqbyrggknstwsgesktlvlsnlsysat-COOH 160. H₂N-kgqnqbyrggknstwsgesktlvlsnlsysat-CONH₂ 161. H₂N-kgqnqdyrggknstwsgbsktlvlsnlsysat-COOH 161. H₂N-kgqnqdyrggknstwsgbsktlvlsnlsysat-CONH₂ 162. H₂N-kgqnqbyrggknstwsgbsktlvlsnlsysat-COOH 162 H₂N-kgqnqbyrggknstwsgbsktlvlsnlsysat-CONH₂ 163. H₂N-kgbnqdyrlgknstwsgbsktlvlsnlsysat-COOH 163. H₂N-kgbnqdyrlgknstwsgbsktlvlsnlsysat-CONH₂ 164. H₂N-kgbnqdyrlgknstwsgbskt-COOH 164. H₂N-kgbnqdyrlgknstwsgbskt-CONH₂ 131. H₂N-katfikvpqnqngkskgyafi-COOH 131. H₂N-katfikvpqnqngkskgyafi-CONH₂ 165. H₂N-katfikvpqnqnxkskgyafi-COOH 165. H₂N-katfikvpqnqnxkskgyafi-CONH₂ 166. H₂N-katfikvpqnqnlkskgyafi-COOH 166. H₂N-katfikvpqnqnlkskgyafi-CONH₂ 167. H₂N-katfikvpqnqnykskgyafi-COOH 167. H₂N-katfikvpqnqnykskgyafi-CONH₂ 168. H₂N-katfikvpqnqnfkskgyafi-COOH 168. H₂N-katfikvpqnqnfkskgyafi-CONH₂ 169. H₂N-katfikvpqnqnwkskgyafi-COOH 169. H₂N-katfikvpqnqnwkskgyafi-CONH₂ 170. H₂N-katfikvpqnqngkskgy-COOH 170. H₂N-katfikvpqnqngkskgy-CONH₂ 171. H₂N-katfikvpqnqnxkskgy-COOH 171. H₂N-katfikvpqnqnxkskgy-CONH₂ 172. H₂N-katfikvpqnqnlkskgy-COOH 172. H₂N-katfikvpqnqnlkskgy-CONH₂ 173. H₂N-katfikvpqnqnykskgy-COOH 173. H₂N-katfikvpqnqnykskgy-CONH₂ 174. H₂N-katfikvpqnqnfkskgy-COOH 174. H₂N-katfikvpqnqnfkskgy-CONH₂ 175. H₂N-katfikvpqnqnwkskgy-COOH 175. H₂N-katfikvpqnqnwkskgy-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00012 TABLE 12 Engineered peptides based on Csp3 protein (caspase 3; apoptosis effector/nuclear encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 176. ¹⁰sksiknlepkiihgs²⁴ 177. .sup.136lkkitnffrgdrcrsltgkpklfiiqacrgt¹⁶⁶ 178. .sup.215fiqslcamlkqyadklefmhiltrvnrkvat²⁴⁵ Synthetic Peptides 176. H₂N-sksiknlepkiihgs-COOH 176. H₂N-sksiknlepkiihgs-CONH₂ 179. H₂N-sksiknlbpkiihgs-COOH 179. H₂N-sksiknlbpkiihgs-CONH₂ 180. H₂N-sksiknlepkiiygs-COOH 180. H₂N-sksiknlepkiiygs-CONH₂ 181. H₂N-sksiknlepkiiybs-COOH 181. H₂N-sksiknlepkiiybs-CONH₂ 182. H₂N-sksiknlbpkiiybs-COOH 182. H₂N-sksiknlbpkiiybs-CONH₂ 177. H₂N-lkkitnffrgdrcrsltgkpklfiiqacrgt-COOH 177. H₂N-lkkitnffrgdrcrsltgkpklfiiqacrgt-CONH₂ 183. H₂N-lkkitnffrgbrcrsltgkpklfiiqacrgt-COOH 183. H₂N-lkkitnffrgbrcrsltgkpklfiiqacrgt-CONH₂ 184. H₂N-lkkitnffrgdrxrsltgkpklfiiqacrgt-COOH 184. H₂N-lkkitnffrgdrxrsltgkpklfiiqacrgt-CONH₂ 185. H₂N-lkkitnffrgdrcrsltgkpklfiiqaxrgt-COOH 185. H₂N-lkkitnffrgdrcrsltgkpklfiiqaxrgt-CONH₂ 186. H₂N-lkkitnfrgbrxrsltgkpklfiiqaxrgt-COOH 186. H₂N-lkkitnfrgbrxrsltgkpklfiiqaxrgt-CONH₂ 187. H₂N-lkkitnfrgbrxrsltgk-COOH 187. H₂N-lkkitnfrgbrxrsltgk-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00013 TABLE 13 Engineered peptides based on Bad protein (apoptotic protein/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 188. ⁹³frgrsrsappnlwaaqrygrelrr¹¹⁶ 189. ¹¹5rrmsdefvdsfkkglprpksagtatq¹⁴⁰ 190. ¹²1fvdsfkkglprpksagtatq¹⁴⁰ Synthetic Peptides 188. H₂N-frgrsrsappnlwaaqrygrelrr-COOH 188. H₂N-frgrsrsappnlwaaqrygrelrr-CONH₂ 191. H₂N-frgrsrsappnlwaaqrygrblrr-COOH 191. H₂N-frgrsrsappnlwaaqrygrblrr-CONH₂ 189. H₂N-rrmsdefvdsfkkglprpksagtatq-COOH 189. H₂N-rrmsdefvdsfkkglprpksagtatq-CONH₂ 192. H₂N-rrmsbefvdsfkkglprpksagtatq-COOH 192. H₂N-rrmsbefvdsfkkglprpksagtatq-CONH₂ 193. H₂N-rrmsdbfvdsfkkglprpksagtatq-COOH 193. H₂N-rrmsdbfvdsfkkglprpksagtatq-CONH₂ 194. H₂N-rrmsdefvbsfkkglprpksagtatq-COOH 194. H₂N-rrmsdefvbsfkkglprpksagtatq-CONH₂ 195. H₂N-rrmsbbfvbsfkkglprpksagtatq-COOH 195. H₂N-rrmsbbfvbsfkkglprpksagtatq-CONH₂ 196. H₂N-rrxsbbfvbsfkkglprpksagtatq-COOH 196. H₂N-rrxsbbfvbsfkkglprpksagtatq-CONH₂ 190. H₂N-fvdsfkkglprpksagtatq-COOH 190. H₂N-fvdsfkkglprpksagtatq-CONH₂ 197. H₂N-fvbsfkkglprpksagtatq-COOH 197. H₂N-fvbsfkkglprpksagtatq-CONH₂ 198. H₂N-fvbsfkkglxrpksag-COOH 198. H₂N-fvbsfkkglxrpksag-CONH₂ 199. H₂N-fvbsfkkglyrpksag-COOH 199. H₂N-fvbsfkkglyrpksag-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00014 TABLE 14 Engineered peptides based on Prf-1 protein (perforin-1/apoptotic/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 200. ³²krshkfvpgawlag⁴⁵ 201. ⁴⁹vtslrrsgsfpvdtqrflr⁶⁸ 202. ¹²3rsirndwkvgldvtpk¹³⁸ 203. ³56rrealrralsqyltdrarwr³75 204. ⁵20nlnhghlkfryhar⁵34 Synthetic Peptides 200. H₂N-krshkfvpgawlag-COOH 200. H₂N-krshkfvpgawlag-CONH₂ 205. H₂N-krshkfvxgawlag-COOH 205. H₂N-krshkfvxgawlag-CONH₂ 201. H₂N-vtslrrsgsfpvdtqrflr-COOH 201. H₂N-vtslrrsgsfpvdtqrflr-CONH₂ 206. H₂N-vtslrrsgsfxvdtqrflr-COOH 206. H₂N-vtslrrsgsfxvdtqrflr-CONH₂ 207. H₂N-vtslrrsgsfpvbtqrflr-COOH 207. H₂N-vtslrrsgsfpvbtqrflr-CONH₂ 208. H₂N-vtslrrsgsfxvbtqrflr-COOH 208. H₂N-vtslrrsgsfxvbtqrflr-CONH₂ 202. H₂N-rsirndwkvgldvtpk-COOH 202. H₂N-rsirndwkvgldvtpk-CONH₂ 209. H₂N-rsirnbwkvgldvtpk-COOH 209. H₂N-rsirnbwkvgldvtpk-CONH₂ 210. H₂N-rsirndwkvgldvt-COOH 210. H₂N-rsirndwkvgldvt-CONH₂ 211. H₂N-rsirnbwkvgldvt-COOH 211. H₂N-rsirnbwkvgldvt-CONH₂ 203. H₂N-rrealrralsqyltdrarwr-COOH 203. H₂N-rrealrralsqyltdrarwr-CONH₂ 212. H₂N-rrbalrralsqyltdrarwr-COOH 212. H₂N-rrbalrralsqyltdrarwr-CONH₂ 213. H₂N-rrealrralsqyltbrarwr-COOH 213. H₂N-rrealrralsqyltbrarwr-CONH₂ 214. H₂N-rrbalrralsqyltbrarwr-COOH 214. H₂N-rrbalrralsqyltbrarwr-CONH₂ 215. H₂N-rxbalrralsqyltbrarwr-COOH 215. H₂N-rxbalrralsqyltbrarwr-CONH₂ 204 H₂N-nlnhghlkfryhar-COOH 204. H₂N-nlnhghlkfryhar-CONH₂ 216. H₂N-nlnbghlkfryhar-COOH 216. H₂N-nlnbghlkfryhar-CONH₂ 217. H₂N-nlnbgblkfryhar-COOH 217. H₂N-nlnbgblkfryhar-CONH₂ 218. H₂N-nlnbgblkfrybar-COOH 218. H₂N-nlnbgblkfrybar-CONH₂ 219. H₂N-nlnxghlkfryhar-COOH 219. H₂N-nlnxghlkfryhar-CONH₂ 220. H₂N-nlnbgxlkfryhar-COOH 220. H₂N-nlnbgxlkfryhar-CONH₂ 221. H₂N-nlnbgblkfryxar-COOH 221. H₂N-nlnbgblkfryxar-CONH₂ 222. H₂N-olnbgblkfrybar-COOH 222. H₂N-olnbgblkfrybar-CONH₂ 223. H₂N-olnbgblkfryxar-COOH 223. H₂N-olnbgblkfryxar-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue; o = anthrylalanine or other non-natural amino acid.

TABLE-US-00015 TABLE 15 Engineered peptides based on Granulysin protein (granulysin-1/apoptotic/human/nuclear-encoded). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 224. ⁶²lgrdyrtcltivqklkk⁷⁸ 225. ⁸²kptqrsvsnaatrvcrtgrsrwr¹⁰4 226. ¹⁰1srwrrryqsrvtqglvag¹²⁵ Synthetic Peptides 224. H₂N-lgrdyrtcltivqklkk-COOH 224. H₂N-lgrdyrtcltivqklkk-CONH₂ 227. H₂N-lgrbyrtcltivqklkk-COOH 227. H₂N-lgrbyrtcltivqklkk-CONH₂ 228. H₂N-lgrdyrtxltivqklkk-COOH 228. H₂N-lgrdyrtxltivqklkk-CONH₂ 229. H₂N-lgrbyrtxltivqklkk-COOH 229. H₂N-lgrbyrtxltivqklkk-CONH₂ 225. H₂N-kptqrsvsnaatrvcrtgrsrwr-COOH 225. H₂N-kptqrsvsnaatrvcrtgrsrwr-CONH₂ 230. H₂N-kptqrsvsnaatrvxrtgrsrwr-COOH 230. H₂N-kptqrsvsnaatrvxrtgrsrwr-CONH₂ 231. H₂N-kptqrsvsnaatrvxrtg-COOH 231. H₂N-kptqrsvsnaatrvxrtg-CONH₂ 232. H₂N-kptqrsvsnyatrvxrtg-COOH 232. H₂N-kptqrsvsnyatrvxrtg-CONH₂ 233. H₂N-kptqrsvsnfatrvxrtg-COOH 233. H₂N-kptqrsvsnfatrvxrtg-CONH₂ 226. H₂N-srwrrryqsrvtqglvag-COOH 226. H₂N-srwrrryqsrvtqglvag-CONH₂ 234. H₂N-srwrryqsrvtqylvag-COOH 234. H₂N-srwrryqsrvtqylvag-CONH₂ 235. H₂N-orwrryqsrvtqylvag-COOH 235. H₂N-orwrryqsrvtqylvag-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00016 TABLE 16 Engineered peptides based on CidA protein (pro-programmed cell death protein/S. aureus). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 236. ²⁸qkifhlplagsivglflfylllqfkiv⁵⁴ 237. ⁸8eitlnyilffaviiigtcivalssgyiaekmsvkhkqrkgi¹²⁷ Synthetic Peptides 236. H₂N-qkifhlplagsivglflfylllqfkiv-COOH 236. H₂N-qkifhlplagsivglflfylllqfkiv-CONH₂ 238. H₂N-qkifhlplabsivglflfylllqfkiv-COOH 238. H₂N-qkifhlplabsivglflfylllqfkiv-CONH₂ 239. H₂N-qkifhlplagsivglflfylglqfkiv-COOH 239. H₂N-qkifhlplagsivglflfylglqfkiv-CONH₂ 240. H₂N-qkifhlplabsivglflfylglqfkiv-COOH 240. H₂N-qkifhlplabsivglflfylglqfkiv-CONH₂ 241. H₂N-labsivglflfylglqfkiv-COOH 241. H₂N-labsivglflfylglqfkiv-CONH₂ 242. H₂N-labsivblflfylglqfkiv-COOH 242. H₂N-labsivblflfylglqfkiv-CONH₂ 237. H₂N-eitlnyilffaviiigtcivalssgyiaekmsvkhkqrkgi-COOH 237. H₂N-eitlnyilffaviiigtcivalssgyiaekmsvkhkqrkgi-CONH₂ 243. H₂N-eitlnyilffaviiigtxivalssgyiaekxsvkhkqrkgi-COOH 243. H₂N-eitlnyilffaviiigtxivalssgyiaekxsvkhkqrkgi-CONH₂ 244. H₂N-aekmsvkhkqrkgi-COOH 244. H₂N-aekmsvkhkqrkgi-CONH₂ 245. H₂N-abkmsvkhkqrkgi-COOH 245. H₂N-abkmsvkhkqrkgi-CONH₂ 246. H₂N-alkmsvkhkqrkgi-COOH 246. H₂N-alkmsvkhkqrkgi-CONH₂ 247. H₂N-alkxsvkhkqrkgi-COOH 247. H₂N-alkxsvkhkqrkgi-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00017 TABLE 17 Engineered peptides based on LrgA protein (anti-programmed cell death protein/S. aureus). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 248. ¹²3kvtsrskgdkvtkkiki¹³⁹ Synthetic Peptides 248. H₂N-kvtsrskgdkvtkkiki-COOH 248. H₂N-kvtsrskgdkvtkkiki-CONH₂ 249. H₂N-kvtsrskgdkvtkwiki-COOH 249. H₂N-kvtsrskgdkvtkwiki-CONH₂ 250. H₂N-kvtsrskgdkvtkziki-COOH 250. H₂N-kvtsrskgdkvtkziki-CONH₂ 251. H₂N-kvtsrskgdkvtkxiki-COOH 251. H₂N-kvtsrskgdkvtkxiki-CONH₂ Underlined residues indicate substitutions; z = d, e, other anionic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00018 TABLE 18 Engineered peptides based on Lambda S21 protein (lytic regulator protein/λ21 phage). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 252. ⁴⁴slvlgfltyltnlyfkiredrrkaarge⁷¹ Synthetic Peptides 252. H₂N-slvlgfltyltnlyfkiredrrkaarge-COOH 252. H₂N-slvlgfltyltnlyfkiredrrkaarge-CONH₂ 253. H₂N-slvlgfltyltnlyfkirbdrrkaarge-COOH 253. H₂N-slvlgfltyltnlyfkirbdrrkaarge-CONH₂ 254. H₂N-slvlgfltyltnlyfkirebrrkaarge-COOH 254. H₂N-slvlgfltyltnlyfkirebrrkaarge-CONH₂ 255. H₂N-slvlgfltyltnlyfkirxxrrkaarge-COOH 255. H₂N-slvlgfltyltnlyfkirxxrrkaarge-CONH₂ 256. H₂N-lyfkirxxrrkaarg-COOH 256. H₂N-lyfkirxxrrkaarg-CONH₂ Underlined residues indicate substitutions; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00019 TABLE 19 Engineered peptides based on Holin protein (lytic regulatory protein/Enterobacteria λ phage). SEQ ID NO: Amino Acid Sequence Native Sequence Domains 257. ²⁷aylrgrynggaftktvi⁴⁴ 258. ⁸⁴sigslikrfaakkagvedgrnq¹⁰5 259. ⁸⁴sigslikrfaakkagv¹⁰0 Synthetic Peptides 257. H₂N-aylrgrynggaftktvi-COOH 257. H₂N-aylrgrynggaftktvi-CONH₂ 260. H₂N-oylrgrynggaftktvi-COOH 260. H₂N-oylrgrynggaftktvi-CONH₂ 258. H₂N-sigslikrfaakkagvedgrnq-COOH 258. H₂N-sigslikrfaakkagvedgrnq-CONH₂ 261. H₂N-sigslikrfaakkagvbdgrnq-COOH 261. H₂N-sigslikrfaakkagvbdgrnq-CONH₂ 262. H₂N-sigslikrfaakkagvebgrnq-COOH 262. H₂N-sigslikrfaakkagvebgrnq-CONH₂ 259 H₂N-sigslikrfaakkagv-COOH 259. H₂N-sigslikrfaakkagv-CONH₂ 263. H₂N-sigslikrfaxkkagv-COOH 263. H₂N-sigslikrfaxkkagv-CONH₂ Underlined residues indicate substitutions; o = anthrylalanine or other non-natural amino acid; b = k, r, n, q, other natural/non-natural basic residue; x = s, t, y, other natural/non-natural H-bond donor/acceptor residue.

TABLE-US-00020 TABLE 20 Additional engineered peptides based on programmed cell death effector proteins. Native SEQ ID Protein NO: Amino Acid Sequence Human 288 H₂N-SQSNRELVVDFLSYKLSQK-COOH Bcl-xL Human 288 H₂N-SQSNRELVVDFLSYKLSQK-CONH₂ Bcl-xL Human 289 H₂N-QKLKKMVDKPTQRSVSN-COOH CTL Granulysin Human 289 H₂N-QKLKKMVDKPTQRSVSN-CONH₂ CTL Granulysin

TABLE-US-00021 TABLE 21 Novel Therapeutic Peptide Designs Based on Programmed Cell Death Effector Domains. ID Name Sequence SEQ ID NO: Length Priority BaxP-I-18 H₂N-alfyfasklvlkalytkv-CONH₂ 264. 18 1 CidA-II-12 H₂N-alkysvkhkqrkgi-CONH₂ 265. 14 2 Ncl-VIII-6 H₂N-irlvskygkskgiayi-CONH₂ 152. 16 3 Csp3-II-12 H₂N-lkkitnfrgkryrsltgk-CONH₂ 266. 18 4 Dnm2-II-4 H₂N-alrsklqsqllslrk-CONH₂ 267. 15 5 Dnm1-IV-2 H₂N-atvkkqvqklk-CONH₂ 80. 11 6 BclXb-I-2 H₂N-flsyklsqkgyswsqfs-CONH₂ 26. 17 7 Hol-III-4 H₂N-sigslikrfaykkagv-CONH₂ 268. 16 8 Mfn1-II-2 H₂N-kiqnnskllrnkavql-CONH₂ 64. 16 9 BclWP-I-4 H₂N-tralvakfvgyklrqkgyv-CONH₂ 269. 19 10 LrgA-I-4 H₂N-kvtsrskgdkvtkwiki-CONH₂ 249. 17 11 BaxP-I-1 H₂N-nfnwgrvvalfyfasklvlkalytkv-CONH₂ 270. 26 BaxP-II-8 H₂N-tvtifvakvltasltiwkk-CONH₂ 271. 19 BclWP-II-6 H₂N-trfrrtfsklaaqlhvt-CONH₂ 272. 17 BclXb-IV-10 H₂N-gqrsptalslylfllywvivk-CONH₂ 273. 21 Mfn1-II-20 H₂N-kkievlkslqskakllrnkagwl-CONH₂ 274. 23 Dnm1-III-4 H₂N-glrnklqsqllsikk-CONH₂ 275. 15 Dnm2-I-4 H₂N-klakkvdpqglrtigvitkl-CONH₂ 276. 21 Dnm2-VI-16 H₂N-kskkytlplknlkir-CONH₂ 277. 15 Csp3-I-10 H₂N-sksiknlkpkiiyks-CONH₂ 278. 15 CidA-I-12 H₂N-laksivrlflfylglqfkiv-CONH₂ 279. 20

Example II

In Vitro Antimicrobial Assay

[0160] The following assay is designed to measure the relative antimicrobial activity of peptides by determining zones of growth inhibition.

[0161] The top eleven prioritized target sequences identified in Example I (see Table 21) were synthesized by solid-phase chain extension synthesis using conventional techniques. Each synthetic peptide was purified by RP-HPLC, and authenticated for purity and correct sequence by mass spectroscopy. Stock concentrations of the synthetic peptides were prepared at 1 mg/mL in 0.01% acetic acid and adjusted to pH 7.2. Synthetic peptides were assessed for antimicrobial efficacy, spectra, and conditional optima (pH 5.5 or 7.5) using the following modified radial diffusion assay, as detailed in Yount and Yeaman, PNAS 1010:7363-7368 (2004).

[0162] Media Preparation

[0163] Molecular grade agarose (1.0%) in 10 mM NaH₂PO₄H₂O was prepared, pH adjusted to 7.5 or 5.5, and autoclaved for 15 minutes at 121° C., then held in a waterbath set at 48° C. until used. Mueller Hinton II overlay agarose was prepared by adding molecular grade agarose to Mueller Hinton II Broth at a final concentration of 1.0%, pH adjusted to 7.5 or 5.5, autoclaved for 10 minutes at 121° C., and then held at 48° C. until used.

[0164] Inoculum Preparation

[0165] Trypticase Soy Broth (TSB) (10 mL) we inoculated with an overnight growth of the test organism and incubated three to six hours until the organism reached log phase. The cells were collected by centrifugation, washed in PBS, then 0.01% acetic acid adjusted to pH 7.2. The pellet was resuspended in TSB and standardized to a 0.5 McFarland turbidity standard. A 10 μl aliquot of the inoculum is added to 10 mL of the pH-adjusted 1.0% molecular grade agarose cooled to 48° C. resulting in a final inoculum concentration of 5×10⁵ CFU/mL. The suspension is poured into a 15×100 mm Petri dish and allowed to solidify.

[0166] After solidification had occurred, five 4 mm diameter wells were bored into the agarose. The central well was used as the acetic acid control while 10 μl of peptide stock solution was added to each of the other wells resulting in a final concentration of 10 μg peptide/well. The plates were incubated upright for three hours at 37° C., then overlaid with 10 mL of Mueller Hinton II agarose. After the overlay solidified, the plates were inverted and incubated overnight at 37° C.

[0167] Activity Determination

[0168] The synthetic peptides identified in Table 22 were assayed for antimicrobial activity against known pathogenic microorganisms. These pathogenic microorganisms included five species of bacteria (Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and Bacillus subtilis) and one species of fungi (Candida albicans) (see Table 23). Zones of growth inhibition were measured and were grouped as complete and/or partial growth zones for the assayed microorganism. Zones were considered complete clearance when there was no visible growth (i.e. completely clear or free of growth). Zones were considered partial clearance when growth was impeded or partially cleared (i.e. reduction in microbial density as compared to adjacent confluent growth). The larger the zone size, the greater the antimicrobial activity of the test peptide. The lack of a zone is an indication of no antimicrobial activity of the test peptide against the target organism under the conditions tested.

TABLE-US-00022 TABLE 22 Peptide key for antimicrobial assay (FIGS. 30-35). Template Identifier* Design Sequence SEQ ID NO: Length Holin protein Hol-III-4 H₂N-sigslikrfaykkagv-CONH₂ 268. 16 Dynamin-2 Dnm2-II-4 H₂N-alrsklqsqllslrk-CONH₂ 267. 15 BclW protein BclWP-I-4 H₂N-tralvakfvgyklrqkgyv-CONH₂ 269. 19 Caspase-3 Csp3-II-12 H₂N-lkkitnfrgkryrsltgk-CONH₂ 266 18 LrgA protein LrgA-I-4 H₂N-kvtsrskgdkvtkwiki-CONH₂ 249. 17 Dynamin-1 Dnm1-IV-2 H₂N-atvkkqvqklk-CONH₂ 80. 11 BclXb protein BclXb-I-2 H₂N-flsyklsqkgyswsqfs-CONH₂ 26. 17 Nucleolin Ncl-VIII-6 H₂N-irlvskygkskgiayi-CONH₂ 152. 16 Mitofusin-1 Mfn1-II-2 H₂N-kiqnnskllrnkavql-CONH₂ 64. 16 Bax protein BaxP-I-18 H₂N-alfyfasklvlkalytkv-CONH₂ 264. 18 CidA protein CidA-II-12 H₂N-alkysvkhkqrkgi-CONH₂ 265. 14 *Note: identifier formula = [Template]-[Model Domain]-[Design No.]

TABLE-US-00023 TABLE 23 Microorganism key for antimicrobial assay (FIGS. 30-35). Genus/Species Identifier Strain Bacteria Staphylococcus aureus SAISP479C ISP479C Escherichia coli ECML-35 ML-35 Salmonella typhimurium ST14028 14028 Pseudomonas aeruginosa PA01 01 Bacillus subtilis BS6633 ATCC 6633 Fungi Candida albicans CA36082S 36082S

Results

[0169] 85% (46/54) of all peptide and microorganism combinations tested at neutral pH showed antimicrobial activity (FIGS. 30, 32 and 34). The majority of peptides also maintained antimicrobial activity under acidic conditions, i.e. pH 5.5 (FIGS. 31, 33 and 35), albeit at a reduced level. The acidic pH reduced the antimicrobial activity of all peptides when tested against Staphylococcus aureus. Additionally, 64% (7/11), 73% (8/11) and 73% (8/11) of the peptides showed lesser antimicrobial activity when tested against Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium, respectively (FIGS. 31, 33 and 35) at pH 5.5 versus pH 7.5. However, the acidic conditions did not appear to affect the activity of peptides Hol-III-4 (SEQ ID NO:268), Ncl-VIII-6 (SEQ ID NO:152) or BaxP-I-18 (SEQ ID NO:264) against Bacillus subtilis, Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa. Alternatively, the antimicrobial activity was increased for 82% (9/11) of the peptides when tested against the fungal pathogen Candida albicans under acidic conditions. Notably, all peptides assayed showed significant and consistently high antimicrobial activity against Bacillus subtilis regardless of the acidity of the agarose media (FIGS. 30-35, lane designation BS6633).

[0170] The above results show that the designed peptides exerted consistent in vitro efficacy against Bacillus. These results supports the concept that these molecules exploit a targetable evolutionary relationship between prokaryotic organisms and eukaryotic mitochondria. Evidence underscoring the bacterium-to-mitochondrial evolution is consistent with this concept (see Herrmann, TRENDS Micro 11(2):74-79 (2003)). For example, proteins that are believed to mediate ion-permeability transition in mitochondria are likely to have evolved from membrane targeting motifs such as helical pre-sequences present in ancestral Gram-positive organisms such as Bacillus, or other prokaryotes (see von Heijne, EMBO Journal 5:1335-1342 (1986)). Such molecules can target mitochondria, are often comprised of 20-60 amino acids, have the potential to form amphipathic α-helices that segregate hydrophobic and hydrophilic facets, and have one facet that is positively charged. The peptides described herein are highly consistent with such molecules. Therefore, the peptides described herein which have antimicrobial activity will also have anti-cancer, anti-inflammatory, anti-rheumatologic and other efficacy by virtue of their likelihood to target mitochondria and induce or cause dysfunctions in programmed cell death circuits.

Example III

Antimicrobial Activity Against Pseudomonas aeruginos and Acinetobacter spp. Strains

[0171] Further to the methods disclosed in Example II, utilizing the same antimicrobial assay above, the antimicrobial activity of peptides Hol-III-4 (SEQ ID NO: 268) and Ncl-VIII-6 (SEQ ID NO: 152 was determined by identifying zones of growth inhibition.

[0172] Hol-III-4 and Ncl-VIII-6 peptides were assayed for antimicrobial activity against a panel of drug-resistant Gram-negative bacterial pathogens, specifically Pseudomonas aeruginosa, and various Acinetobacter spp. strains. The efficacies of these peptides were tested in the context of pH 5.5 (FIGS. 36-46) and pH 7.5 (FIGS. 47-57) conditions, and compared with other peptides known to have antimicrobial activity (e.g. RP-1, 6W-RP-1 (a 6-Trp variant of RP-1), IK, and PMP-2), in the radial diffusion assay. The RP-1 peptide is well known in the art to have antimicrobial activity, as illustrated in Yeaman et al., Antimicrobial Agents and Chemotherapy, 46(12):3883-3891 (2002). The 6W-RP-1 peptide is a 6-Trp variant of RP-1, which also has antimicrobial activity, as illustrated in Kilelee et al, Antimicrobial Agents and Chemotherapy 54(10):4476-4479 (2010). PMP is the C-terminal helix of the consensus molecule cPMP, as shown in Table 1 (bottom row; N-AALYKKKIIKKLLES-C; as shown in Yeaman et al., Bichimica et Biophysica Acta, 1768:609-619 (2007). The IK peptide is designed to have a nearly maximal polar angle (maximum angle is approximately 180°). The results of the antimicrobial assay show that both Hol-III-4 and Ncl-VIII-6 consistently showed significant antimicrobial activity against all Pseudomonas aeruginosa, and all Acinetobacter spp. strains tested. Additionally, the antimicrobial activity of both Hol-III-4 and Ncl-VIII-6 against the various Acinetobacter baumannii isolates tested appeared to be pH dependent (see FIGS. 39, 40, 43-46 vs. FIGS. 50, 51, 54-57)

[0173] These results expand on and further substantiate the results shown in Example II regarding the antimicrobial efficacy of the peptides disclosed here. Additionally, the current data supports the conclusion that the PCD peptides may have a unique mechanism of action, but achieve generally equivalent efficacy as RP-1-like peptides against most organisms tested. The is evident by the following observations. The RP-1 and related antimicrobial peptides appear to target microbial cells enriched with electronegative constituents (e.g. phosphatidylglycerol, cardiolipin, etc.), and/or those having electronegative transmembrane potential. These features, in addition to conformational plasticity, inhibition of intracellular functions (e.g. macromolecular synthesis) and possibly superstructural assembly, are believed to participate in the preferential microbial targeting and antimicrobial effects of such peptides. By comparison, the current peptides are designed from programmed cell death effector or modulating proteins. Without being bound by theory, one hypothesis is that such peptides induce microbial and other target cell death by inducing or dysregulating programmed cell death. Thus, it is possible that these peptides function via a mechanism that is not identical to the RP-1-like peptides. In balance, it is also possible that PCD peptides evolved (e.g. diverged) from antimicrobial peptide sequences, based on the view that mitochondria are modern day "bacteria"; this is one hypothesis we posed in the patent. If so, then the mechanisms may be conserved among antimicrobial helices, even if the helices come from proteins believed to have vastly divergent functions (e.g. helices from PMPs, PCD proteins, chemokines, etc).

[0174] Throughout this application various publications have been referenced. The disclosures of these publications in their entireties are hereby incorporated by reference in this application in order to more fully describe the state of the art to which this invention pertains. Although the invention has been described with reference to the examples provided above, it should be understood that various modifications can be made without departing from the spirit of the invention.

Sequence CWU 1

1

428126PRTArtificial SequenceEngineered peptides based on Bax protein 1Asn Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys1 5 10 15 Leu Val Leu Lys Ala Leu Cys Thr Lys Val 20 25 222PRTArtificial SequenceEngineered peptides based on Bax protein 2Thr Trp Gln Thr Val Thr Ile Phe Val Ala Gly Val Leu Thr Ala Ser1 5 10 15 Leu Thr Ile Trp Lys Lys 20 326PRTArtificial SequenceEngineered peptides based on Bax protein 3Asn Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys1 5 10 15 Leu Val Leu Lys Ala Leu Xaa Thr Lys Val 20 25 426PRTArtificial SequenceEngineered peptides based on Bax protein 4Asn Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys1 5 10 15 Leu Val Leu Lys Ala Leu Xaa Thr Xaa Val 20 25 523PRTArtificial SequenceEngineered peptides based on Bax protein 5Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu1 5 10 15 Lys Ala Leu Cys Thr Lys Val 20 623PRTArtificial SequenceEngineered peptides based on Bax protein 6Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu1 5 10 15 Lys Ala Leu Xaa Thr Lys Val 20 721PRTArtificial SequenceEngineered peptides based on Bax protein 7Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu Lys Ala1 5 10 15 Leu Cys Thr Lys Val 20 821PRTArtificial SequenceEngineered peptides based on Bax protein 8Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu Lys Ala1 5 10 15 Leu Xaa Thr Lys Val 20 918PRTArtificial SequenceEngineered peptides based on Bax protein 9Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu Lys Ala Leu Cys Thr1 5 10 15 Lys Val1018PRTArtificial SequenceEngineered peptides based on Bax protein 10Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu Lys Ala Leu Xaa Thr1 5 10 15 Lys Val1122PRTArtificial SequenceEngineered peptides based on Bax protein 11Thr Trp Gln Thr Val Thr Ile Phe Val Ala Xaa Val Leu Thr Ala Ser1 5 10 15 Leu Thr Ile Trp Lys Lys 20 1219PRTArtificial SequenceEngineered peptides based on Bax protein 12Thr Val Thr Ile Phe Val Ala Gly Val Leu Thr Ala Ser Leu Thr Ile1 5 10 15 Trp Lys Lys1319PRTArtificial SequenceEngineered peptides based on Bax protein 13Thr Val Thr Ile Phe Val Ala Xaa Val Leu Thr Ala Ser Leu Thr Ile1 5 10 15 Trp Lys Lys1419PRTArtificial SequenceEngineered peptides based on Bcl-W protein 14Thr Arg Ala Leu Val Ala Asp Phe Val Gly Tyr Lys Leu Arg Gln Lys1 5 10 15 Gly Tyr Val1517PRTArtificial SequenceEngineered peptides based on Bcl-W protein 15Thr Arg Phe Arg Arg Thr Phe Ser Asp Leu Ala Ala Gln Leu His Val1 5 10 15 Thr1635PRTArtificial SequenceEngineered peptides based on Bcl-W protein 16Ala Arg Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val Leu1 5 10 15 Thr Gly Ala Val Ala Leu Gly Ala Leu Val Thr Val Gly Ala Phe Phe 20 25 30 Ala Ser Lys 35 1719PRTArtificial SequenceEngineered peptides based on Bcl-W protein 17Thr Arg Ala Leu Val Ala Xaa Phe Val Gly Tyr Lys Leu Arg Gln Lys1 5 10 15 Gly Tyr Val1817PRTArtificial SequenceEngineered peptides based on Bcl-W protein 18Thr Arg Phe Arg Xaa Thr Phe Ser Asp Leu Ala Ala Gln Leu His Val1 5 10 15 Thr1917PRTArtificial SequenceEngineered peptides based on Bcl-W protein 19Thr Arg Phe Arg Arg Thr Phe Ser Xaa Leu Ala Ala Gln Leu His Val1 5 10 15 Thr2034PRTArtificial SequenceEngineered peptides based on Bcl-W protein 20Arg Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr1 5 10 15 Gly Ala Val Ala Leu Gly Ala Leu Val Thr Val Gly Ala Phe Phe Ala 20 25 30 Ser Lys2134PRTArtificial SequenceEngineered peptides based on Bcl-W protein 21Arg Arg Leu Arg Xaa Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr1 5 10 15 Gly Ala Val Ala Leu Gly Ala Leu Val Thr Val Gly Ala Phe Phe Ala 20 25 30 Ser Lys2234PRTArtificial SequenceEngineered peptides based on Bcl-W protein 22Arg Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr1 5 10 15 Xaa Ala Val Ala Leu Gly Ala Leu Val Thr Val Gly Ala Phe Phe Ala 20 25 30 Ser Lys2334PRTArtificial SequenceEngineered peptides based on Bcl-W protein 23Arg Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr1 5 10 15 Gly Ala Val Ala Leu Xaa Ala Leu Val Thr Val Gly Ala Phe Phe Ala 20 25 30 Ser Lys2434PRTArtificial SequenceEngineered peptides based on Bcl-W protein 24Arg Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr1 5 10 15 Gly Ala Val Ala Leu Gly Ala Leu Val Thr Val Xaa Ala Phe Phe Ala 20 25 30 Ser Lys2534PRTArtificial SequenceEngineered peptides based on Bcl-W protein 25Arg Arg Leu Arg Xaa Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr1 5 10 15 Xaa Ala Val Ala Leu Xaa Ala Leu Val Thr Val Xaa Ala Phe Phe Ala 20 25 30 Ser Lys2617PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 26Phe Leu Ser Tyr Lys Leu Ser Gln Lys Gly Tyr Ser Trp Ser Gln Phe1 5 10 15 Ser2725PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 27Leu Arg Tyr Arg Arg Ala Phe Ser Asp Leu Thr Ser Gln Leu His Ile1 5 10 15 Thr Pro Gly Thr Ala Tyr Gln Ser Phe 20 25 2820PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 28Gly Trp Val Arg Thr Lys Pro Leu Val Cys Pro Phe Ser Leu Ala Ser1 5 10 15 Gly Gln Arg Ser 20 2921PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 29Gly Gln Arg Ser Pro Thr Ala Leu Leu Leu Tyr Leu Phe Leu Leu Cys1 5 10 15 Trp Val Ile Val Gly 20 3025PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 30Leu Arg Tyr Arg Arg Ala Phe Ser Xaa Leu Thr Ser Gln Leu His Ile1 5 10 15 Thr Pro Gly Thr Ala Tyr Gln Ser Phe 20 25 3120PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 31Gly Trp Val Arg Thr Lys Pro Leu Val Xaa Pro Phe Ser Leu Ala Ser1 5 10 15 Gly Gln Arg Ser 20 3220PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 32Gly Trp Val Arg Thr Lys Pro Leu Val Xaa Pro Phe Ser Leu Ala Ser1 5 10 15 Xaa Gln Arg Ser 20 3321PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 33Gly Gln Arg Ser Pro Thr Ala Leu Xaa Leu Tyr Leu Phe Leu Leu Cys1 5 10 15 Trp Val Ile Val Gly 20 3421PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 34Gly Gln Arg Ser Pro Thr Ala Leu Leu Leu Tyr Leu Phe Leu Leu Xaa1 5 10 15 Trp Val Ile Val Gly 20 3521PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 35Gly Gln Arg Ser Pro Thr Ala Leu Leu Leu Tyr Leu Phe Leu Leu Cys1 5 10 15 Trp Val Ile Val Xaa 20 3621PRTArtificial SequenceEngineered peptides based on Bcl-xBetta protein 36Gly Gln Arg Ser Pro Thr Ala Leu Xaa Leu Tyr Leu Phe Leu Leu Xaa1 5 10 15 Trp Val Ile Val Xaa 20 3718PRTArtificial SequenceEngineered peptides based on Bak protein 37Arg Val Val Ala Leu Leu Gly Phe Gly Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln3824PRTArtificial SequenceEngineered peptides based on Bak protein 38Ile Leu Asn Val Leu Val Val Leu Gly Val Val Leu Leu Gly Gln Phe1 5 10 15 Val Val Arg Arg Phe Phe Lys Ser 20 3918PRTArtificial SequenceEngineered peptides based on Bak protein 39Arg Val Val Ala Leu Leu Xaa Phe Gly Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4018PRTArtificial SequenceEngineered peptides based on Bak protein 40Arg Val Val Ala Leu Leu Gly Phe Xaa Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4118PRTArtificial SequenceEngineered peptides based on Bak protein 41Arg Val Val Ala Leu Leu Xaa Phe Xaa Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4218PRTArtificial SequenceEngineered peptides based on Bak protein 42Arg Val Val Ala Leu Tyr Gly Phe Gly Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4318PRTArtificial SequenceEngineered peptides based on Bak protein 43Arg Val Val Ala Leu Trp Gly Phe Gly Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4418PRTArtificial SequenceEngineered peptides based on Bak protein 44Arg Val Val Ala Leu Tyr Xaa Phe Xaa Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4518PRTArtificial SequenceEngineered peptides based on Bak protein 45Arg Val Val Ala Leu Trp Xaa Phe Xaa Tyr Arg Leu Ala Leu His Val1 5 10 15 Tyr Gln4624PRTArtificial SequenceEngineered peptides based on Bak protein 46Ile Leu Asn Val Leu Val Xaa Leu Gly Val Val Leu Leu Gly Gln Phe1 5 10 15 Val Val Arg Arg Phe Phe Lys Ser 20 4718PRTArtificial SequenceEngineered peptides based on Bak protein 47Ile Leu Asn Val Leu Val Xaa Leu Val Leu Gly Gln Phe Val Arg Phe1 5 10 15 Lys Ser4818PRTArtificial SequenceEngineered peptides based on Bcl-2 protein 48Asn Arg Glu Ile Val Met Lys Tyr Ile His Tyr Lys Leu Ser Gln Arg1 5 10 15 Gly Tyr4918PRTArtificial SequenceEngineered peptides based on Bcl-2 protein 49Asn Arg Xaa Ile Val Met Lys Tyr Ile His Tyr Lys Leu Ser Gln Arg1 5 10 15 Gly Tyr5018PRTArtificial SequenceEngineered peptides based on Bcl-2 protein 50Asn Arg Glu Ile Val Xaa Lys Tyr Ile His Tyr Lys Leu Ser Gln Arg1 5 10 15 Gly Tyr5118PRTArtificial SequenceEngineered peptides based on Bcl-2 protein 51Asn Arg Glu Ile Val Met Lys Tyr Ile Xaa Tyr Lys Leu Ser Gln Arg1 5 10 15 Gly Tyr5218PRTArtificial SequenceEngineered peptides based on Bcl-2 protein 52Asn Arg Xaa Ile Val Xaa Lys Tyr Ile Xaa Tyr Lys Leu Ser Gln Arg1 5 10 15 Gly Tyr5316PRTArtificial SequenceEngineered peptides based on Bcl-2 isoform 1 protein 53Leu Ala Leu Arg Gln Ala Gly Asp Asp Phe Ser Arg Arg Tyr Arg Gly1 5 10 15 5416PRTArtificial SequenceEngineered peptides based on Bcl-2 isoform 1 protein 54Leu Ala Leu Arg Gln Ala Gly Xaa Asp Phe Ser Arg Arg Tyr Arg Gly1 5 10 15 5516PRTArtificial SequenceEngineered peptides based on Bcl-2 isoform 1 protein 55Leu Ala Leu Arg Gln Ala Gly Asp Xaa Phe Ser Arg Arg Tyr Arg Gly1 5 10 15 5616PRTArtificial SequenceEngineered peptides based on Bcl-2 isoform 1 protein 56Leu Ala Leu Arg Gln Ala Gly Xaa Xaa Phe Ser Arg Arg Tyr Arg Gly1 5 10 15 5716PRTArtificial SequenceEngineered peptides based on Bcl-2 isoform 1 protein 57Leu Ala Leu Arg Gln Ala Xaa Xaa Xaa Phe Ser Arg Arg Tyr Arg Gly1 5 10 15 5835PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 58Lys Glu Ile Asp Gln Leu Glu Lys Ile Gln Asn Asn Ser Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Val Gln Leu Glu Asn Glu Leu Glu Asn Phe Thr Lys 20 25 30 Gln Phe Leu 35 5935PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 59Lys Glu Ile Xaa Gln Leu Glu Lys Ile Gln Asn Asn Ser Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Val Gln Leu Glu Asn Glu Leu Glu Asn Phe Thr Lys 20 25 30 Gln Phe Leu 35 6035PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 60Lys Glu Ile Xaa Gln Leu Xaa Lys Ile Gln Asn Asn Ser Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Val Gln Leu Xaa Asn Glu Leu Glu Asn Phe Thr Lys 20 25 30 Gln Phe Leu 35 6135PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 61Lys Glu Ile Xaa Gln Leu Xaa Lys Ile Gln Asn Asn Ser Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Val Gln Leu Xaa Asn Glu Leu Glu Asn Phe Thr Lys 20 25 30 Gln Phe Leu 35 6235PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 62Lys Glu Ile Xaa Gln Leu Xaa Lys Ile Gln Asn Asn Ser Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Val Gln Leu Xaa Asn Xaa Leu Glu Asn Phe Thr Lys 20 25 30 Gln Phe Leu 35 6335PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 63Lys Glu Ile Xaa Gln Leu Xaa Lys Ile Gln Asn Asn Ser Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Val Gln Leu Xaa Asn Xaa Leu Xaa Asn Phe Thr Lys 20 25 30 Gln Phe Leu 35 6416PRTArtificial SequenceEngineered peptides based on Mfn-1 protein 64Lys Ile Gln Asn Asn Ser Lys Leu Leu Arg Asn Lys Ala Val Gln Leu1 5 10 15 6540PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 65Asn Lys Lys Ile Glu Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Asp Ser Glu Leu Asn Met Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg 35 40 6640PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 66Asn Lys Lys Ile Xaa Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Asp Ser Glu Leu Asn Met Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg 35 40 6740PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 67Asn Lys Lys Ile Glu Val Leu Xaa Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Asp Ser Glu Leu Asn Met Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg 35 40 6840PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 68Asn Lys Lys Ile Glu Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Xaa Ser Glu Leu Asn Met Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg 35 40 6940PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 69Asn Lys Lys Ile Glu Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Asp Ser Xaa Leu Asn Met Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg

35 40 7040PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 70Asn Lys Lys Ile Glu Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Asp Ser Glu Leu Asn Xaa Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg 35 40 7140PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 71Asn Lys Lys Ile Xaa Val Leu Xaa Ser Leu Gln Ser Lys Ala Lys Leu1 5 10 15 Leu Arg Asn Lys Ala Gly Trp Leu Xaa Ser Xaa Leu Asn Xaa Phe Thr 20 25 30 His Gln Tyr Leu Gln Pro Ser Arg 35 40 7223PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 72Lys Lys Ile Glu Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Gly Trp Leu 20 7323PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 73Lys Lys Ile Xaa Val Leu Asp Ser Leu Gln Ser Lys Ala Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Gly Trp Leu 20 7423PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 74Lys Lys Ile Glu Val Leu Xaa Ser Leu Gln Ser Lys Ala Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Gly Trp Leu 20 7523PRTArtificial SequenceEngineered peptides based on Mfn-2 protein 75Lys Lys Ile Xaa Val Leu Xaa Ser Leu Gln Ser Lys Ala Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Gly Trp Leu 20 7618PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 76Lys Leu Leu Pro Leu Arg Arg Gly Tyr Ile Gly Val Val Asn Arg Ser1 5 10 15 Gln Lys7714PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 77Arg Lys Phe Phe Leu Ser His Pro Ser Tyr Arg His Leu Ala1 5 10 7815PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 78Gly Leu Arg Asn Lys Leu Gln Ser Gln Leu Leu Ser Ile Glu Lys1 5 10 15 7918PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 79Ala Arg Lys Thr Lys Ala Leu Leu Gln Met Val Gln Gln Phe Ala Val1 5 10 15 Asp Phe8011PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 80Ala Thr Val Lys Lys Gln Val Gln Lys Leu Lys1 5 10 8120PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 81Val Ile Arg Lys Gly Trp Leu Thr Ile Asn Asn Ile Gly Ile Met Lys1 5 10 15 Gly Gly Ser Lys 20 8230PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 82Asn Leu Lys Leu Arg Asp Val Glu Lys Gly Phe Met Ser Ser Lys His1 5 10 15 Ile Phe Ala Leu Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 8317PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 83Lys Ala Ser Phe Leu Arg Ala Gly Val Tyr Pro Glu Arg Val Gly Asp1 5 10 15 Lys8415PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 84Gly Leu Arg Asn Lys Leu Gln Ser Gln Leu Leu Ser Ile Xaa Lys1 5 10 15 8518PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 85Ala Arg Lys Thr Lys Ala Leu Leu Gln Met Val Gln Gln Phe Ala Val1 5 10 15 Xaa Phe8630PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 86Asn Leu Lys Leu Arg Xaa Val Glu Lys Gly Phe Met Ser Ser Lys His1 5 10 15 Ile Phe Ala Leu Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 8730PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 87Asn Leu Lys Leu Arg Asp Val Xaa Lys Gly Phe Met Ser Ser Lys His1 5 10 15 Ile Phe Ala Leu Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 8830PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 88Asn Leu Lys Leu Arg Asp Val Glu Lys Gly Phe Met Ser Ser Lys His1 5 10 15 Ile Phe Ala Leu Phe Asn Thr Xaa Gln Arg Asn Val Tyr Lys 20 25 30 8930PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 89Asn Leu Lys Leu Arg Xaa Val Xaa Lys Gly Phe Met Ser Ser Lys His1 5 10 15 Ile Phe Ala Leu Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 9030PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 90Asn Leu Lys Leu Arg Xaa Val Xaa Lys Gly Phe Met Ser Ser Lys His1 5 10 15 Ile Phe Ala Leu Phe Asn Thr Xaa Gln Arg Asn Val Tyr Lys 20 25 30 9117PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 91Lys Ala Ser Phe Leu Arg Ala Gly Val Tyr Pro Xaa Arg Val Gly Asp1 5 10 15 Lys9217PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 92Lys Ala Ser Phe Leu Arg Ala Gly Val Tyr Pro Glu Arg Val Gly Xaa1 5 10 15 Lys9317PRTArtificial SequenceEngineered peptides based on Dnm-1 protein 93Lys Ala Ser Phe Leu Arg Ala Gly Val Tyr Pro Xaa Arg Val Gly Xaa1 5 10 15 Lys9420PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 94Lys Leu Ala Lys Glu Val Asp Pro Gln Gly Leu Arg Thr Ile Gly Val1 5 10 15 Ile Thr Lys Leu 20 9515PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 95Ala Leu Arg Ser Lys Leu Gln Ser Gln Leu Leu Ser Leu Glu Lys1 5 10 15 9616PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 96Thr Arg Lys Thr Lys Ala Leu Leu Gln Met Val Gln Gln Phe Gly Val1 5 10 15 9711PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 97Ala Ile Val Lys Lys Gln Val Val Lys Leu Lys1 5 10 9818PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 98Ala Gln Gln Arg Ser Thr Gln Leu Asn Lys Lys Arg Ala Ile Pro Asn1 5 10 15 Gln Gly9940PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 99Lys Glu Lys Lys Tyr Met Leu Pro Leu Asp Asn Leu Lys Ile Arg Asp1 5 10 15 Val Glu Lys Gly Phe Met Ser Asn Lys His Val Phe Ala Ile Phe Asn 20 25 30 Thr Glu Gln Arg Asn Val Tyr Lys 35 40 10030PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 100Asn Leu Lys Ile Arg Asp Val Glu Lys Gly Phe Met Ser Asn Lys His1 5 10 15 Val Phe Ala Ile Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 10118PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 101Ser Trp Lys Ala Ser Phe Leu Arg Ala Gly Val Tyr Pro Glu Lys Asp1 5 10 15 Gln Ala10220PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 102Lys Leu Ala Lys Xaa Val Asp Pro Gln Gly Leu Arg Thr Ile Gly Val1 5 10 15 Ile Thr Lys Leu 20 10320PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 103Lys Leu Ala Lys Glu Val Xaa Pro Gln Gly Leu Arg Thr Ile Gly Val1 5 10 15 Ile Thr Lys Leu 20 10420PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 104Lys Leu Ala Lys Xaa Val Xaa Pro Gln Gly Leu Arg Thr Ile Gly Val1 5 10 15 Ile Thr Lys Leu 20 10515PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 105Ala Leu Arg Ser Lys Leu Gln Ser Gln Leu Leu Ser Leu Xaa Lys1 5 10 15 10611PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 106Xaa Ile Val Lys Lys Gln Val Val Lys Leu Lys1 5 10 10715PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 107Lys Glu Lys Lys Tyr Met Leu Pro Leu Asp Asn Leu Lys Ile Arg1 5 10 15 10815PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 108Lys Xaa Lys Lys Tyr Met Leu Pro Leu Asp Asn Leu Lys Ile Arg1 5 10 15 10915PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 109Lys Glu Lys Lys Tyr Xaa Leu Pro Leu Asp Asn Leu Lys Ile Arg1 5 10 15 11015PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 110Lys Glu Lys Lys Tyr Met Leu Pro Leu Xaa Asn Leu Lys Ile Arg1 5 10 15 11115PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 111Lys Glu Lys Lys Tyr Met Leu Pro Leu Xaa Asn Leu Lys Ile Arg1 5 10 15 11215PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 112Lys Xaa Lys Lys Tyr Met Leu Pro Leu Xaa Asn Leu Lys Ile Arg1 5 10 15 11315PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 113Lys Xaa Lys Lys Tyr Xaa Leu Pro Leu Xaa Asn Leu Lys Ile Arg1 5 10 15 11422PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 114Lys Gly Phe Met Ser Asn Lys His Val Phe Ala Ile Phe Asn Thr Glu1 5 10 15 Gln Arg Asn Val Tyr Lys 20 11540PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 115Lys Xaa Lys Lys Tyr Met Leu Pro Leu Asp Asn Leu Lys Ile Arg Asp1 5 10 15 Val Glu Lys Gly Phe Met Ser Asn Lys His Val Phe Ala Ile Phe Asn 20 25 30 Thr Glu Gln Arg Asn Val Tyr Lys 35 40 11640PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 116Lys Glu Lys Lys Tyr Xaa Leu Pro Leu Asp Asn Leu Lys Ile Arg Asp1 5 10 15 Val Glu Lys Gly Phe Met Ser Asn Lys His Val Phe Ala Ile Phe Asn 20 25 30 Thr Glu Gln Arg Asn Val Tyr Lys 35 40 11740PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 117Lys Glu Lys Lys Tyr Met Leu Pro Leu Asp Asn Leu Lys Ile Arg Asp1 5 10 15 Val Glu Lys Gly Phe Xaa Ser Asn Lys His Val Phe Ala Ile Phe Asn 20 25 30 Thr Glu Gln Arg Asn Val Tyr Lys 35 40 11830PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 118Asn Leu Lys Ile Arg Xaa Val Glu Lys Gly Phe Met Ser Asn Lys His1 5 10 15 Val Phe Ala Ile Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 11930PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 119Asn Leu Lys Ile Arg Asp Val Xaa Lys Gly Phe Met Ser Asn Lys His1 5 10 15 Val Phe Ala Ile Phe Asn Thr Glu Gln Arg Asn Val Tyr Lys 20 25 30 12030PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 120Asn Leu Lys Ile Arg Xaa Val Xaa Lys Gly Phe Met Ser Asn Lys His1 5 10 15 Val Phe Ala Ile Phe Asn Thr Xaa Gln Arg Asn Val Tyr Lys 20 25 30 12118PRTArtificial SequenceEngineered peptides based on Dnm-2 protein 121Ser Trp Lys Ala Ser Phe Leu Arg Ala Gly Val Tyr Pro Xaa Lys Asp1 5 10 15 Gln Ala12218PRTArtificial SequenceEngineered peptides based on Ncl protein 122Gln Lys Lys Gly Lys Lys Ala Ala Ala Thr Ser Ala Lys Lys Val Val1 5 10 15 Val Ser12316PRTArtificial SequenceEngineered peptides based on Ncl protein 123Thr Lys Lys Val Ala Val Ala Thr Pro Ala Lys Lys Ala Ala Val Thr1 5 10 15 12419PRTArtificial SequenceEngineered peptides based on Ncl protein 124Lys Lys Gly Ala Ala Ile Pro Ala Lys Gly Ala Lys Asn Gly Lys Asn1 5 10 15 Ala Lys Lys12518PRTArtificial SequenceEngineered peptides based on Ncl protein 125Ala Lys Gly Lys Lys Ala Ala Lys Val Val Pro Val Lys Ala Lys Asn1 5 10 15 Val Ala12618PRTArtificial SequenceEngineered peptides based on Ncl protein 126Val Lys Glu Ala Pro Gly Lys Arg Lys Lys Glu Met Ala Lys Gln Lys1 5 10 15 Ala Ala12719PRTArtificial SequenceEngineered peptides based on Ncl protein 127Lys Ala Leu Glu Leu Thr Gly Leu Lys Val Phe Gly Asn Glu Ile Lys1 5 10 15 Leu Glu Lys12825PRTArtificial SequenceEngineered peptides based on Ncl protein 128Lys Gly Lys Asp Ser Lys Lys Glu Arg Asp Ala Arg Thr Leu Leu Ala1 5 10 15 Lys Asn Leu Pro Tyr Lys Val Thr Gln 20 25 12916PRTArtificial SequenceEngineered peptides based on Ncl protein 129Ile Arg Leu Val Ser Lys Asp Gly Lys Ser Lys Gly Ile Ala Tyr Ile1 5 10 15 13032PRTArtificial SequenceEngineered peptides based on Ncl protein 130Lys Gly Gln Asn Gln Asp Tyr Arg Gly Gly Lys Asn Ser Thr Trp Ser1 5 10 15 Gly Glu Ser Lys Thr Leu Val Leu Ser Asn Leu Ser Tyr Ser Ala Thr 20 25 30 13121PRTArtificial SequenceEngineered peptides based on Ncl protein 131Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Gly Lys Ser Lys1 5 10 15 Gly Tyr Ala Phe Ile 20 13218PRTArtificial SequenceEngineered peptides based on Ncl protein 132Gln Lys Lys Gly Lys Lys Ala Xaa Ala Thr Ser Ala Lys Lys Val Tyr1 5 10 15 Val Ser13318PRTArtificial SequenceEngineered peptides based on Ncl protein 133Gln Lys Lys Gly Lys Lys Ala Ala Ala Thr Ser Ala Lys Lys Val Tyr1 5 10 15 Val Ser13418PRTArtificial SequenceEngineered peptides based on Ncl protein 134Gln Lys Lys Gly Lys Lys Ala Ala Ala Thr Ser Ala Lys Lys Val Trp1 5 10 15 Val Ser13518PRTArtificial SequenceEngineered peptides based on Ncl protein 135Gln Lys Lys Gly Lys Lys Ala Xaa Ala Thr Ser Ala Lys Lys Val Tyr1 5 10 15 Val Ser13618PRTArtificial SequenceEngineered peptides based on Ncl protein 136Gln Lys Lys Gly Lys Lys Ala Xaa Ala Thr Ser Ala Lys Lys Val Trp1 5 10 15 Val Ser13719PRTArtificial SequenceEngineered peptides based on Ncl protein 137Lys Lys Gly Ala Xaa Ile Pro Ala Lys Gly Ala Lys Asn Gly Lys Asn1 5 10 15 Ala Lys Lys13818PRTArtificial SequenceEngineered peptides based on Ncl protein 138Ala Lys Gly Lys Lys Ala Ala Lys Val Val Xaa Val Lys Ala Lys Asn1 5 10 15 Val Ala13918PRTArtificial SequenceEngineered peptides based on Ncl protein 139Val Lys Xaa Ala Pro Gly Lys Arg Lys Lys Glu Met Ala Lys Gln Lys1 5 10 15 Ala Ala14018PRTArtificial SequenceEngineered peptides based on Ncl protein 140Val Lys Glu Ala Pro Gly Lys Arg Lys Lys Xaa Met Ala Lys Gln Lys1 5 10 15 Ala Ala14118PRTArtificial SequenceEngineered peptides based on Ncl protein 141Val Lys Glu Ala Pro Gly Lys Arg Lys Lys Glu Xaa Ala Lys Gln Lys1 5 10 15 Ala Ala14218PRTArtificial SequenceEngineered peptides based on Ncl protein 142Val Lys Xaa Ala Pro Gly Lys Arg Lys Lys Xaa Xaa Ala Lys Gln Lys1 5 10 15 Ala Ala14319PRTArtificial SequenceEngineered peptides based on Ncl protein 143Lys Ala Leu Xaa Leu Thr Gly Leu Lys Val Phe Gly Asn Glu Ile Lys1 5 10 15 Leu Glu Lys14419PRTArtificial SequenceEngineered peptides based on Ncl protein 144Lys Ala Leu Glu Leu Thr Gly Leu Lys Val Phe Gly Asn Xaa Ile Lys1 5 10 15 Leu Glu Lys14519PRTArtificial SequenceEngineered peptides based on Ncl protein 145Lys Ala Leu Glu Leu Thr Gly Leu Lys Val Phe Gly Asn Glu Ile Lys1

5 10 15 Leu Xaa Lys14619PRTArtificial SequenceEngineered peptides based on Ncl protein 146Lys Ala Leu Xaa Leu Thr Gly Leu Lys Val Phe Gly Asn Xaa Ile Lys1 5 10 15 Leu Xaa Lys14725PRTArtificial SequenceEngineered peptides based on Ncl protein 147Lys Gly Lys Xaa Ser Lys Lys Glu Arg Asp Ala Arg Thr Leu Leu Ala1 5 10 15 Lys Asn Leu Pro Tyr Lys Val Thr Gln 20 25 14825PRTArtificial SequenceEngineered peptides based on Ncl protein 148Lys Gly Lys Asp Ser Lys Lys Xaa Arg Asp Ala Arg Thr Leu Leu Ala1 5 10 15 Lys Asn Leu Pro Tyr Lys Val Thr Gln 20 25 14925PRTArtificial SequenceEngineered peptides based on Ncl protein 149Lys Gly Lys Asp Ser Lys Lys Glu Arg Xaa Ala Arg Thr Leu Leu Ala1 5 10 15 Lys Asn Leu Pro Tyr Lys Val Thr Gln 20 25 15025PRTArtificial SequenceEngineered peptides based on Ncl protein 150Lys Gly Lys Xaa Ser Lys Lys Xaa Arg Xaa Ala Arg Thr Leu Leu Ala1 5 10 15 Lys Asn Leu Pro Tyr Lys Val Thr Gln 20 25 15116PRTArtificial SequenceEngineered peptides based on Ncl protein 151Ile Arg Leu Val Ser Lys Phe Gly Lys Ser Lys Gly Ile Ala Tyr Ile1 5 10 15 15216PRTArtificial SequenceEngineered peptides based on Ncl protein 152Ile Arg Leu Val Ser Lys Tyr Gly Lys Ser Lys Gly Ile Ala Tyr Ile1 5 10 15 15316PRTArtificial SequenceEngineered peptides based on Ncl protein 153Ile Arg Leu Val Ser Lys Trp Gly Lys Ser Lys Gly Ile Ala Tyr Ile1 5 10 15 15417PRTArtificial SequenceEngineered peptides based on Ncl protein 154Ile Arg Leu Val Ser Lys Leu Trp Gly Lys Ser Lys Gly Ile Ala Tyr1 5 10 15 Ile15512PRTArtificial SequenceEngineered peptides based on Ncl protein 155Ile Arg Leu Val Ser Lys Asp Gly Lys Ser Lys Gly1 5 10 15613PRTArtificial SequenceEngineered peptides based on Ncl protein 156Ile Arg Leu Val Ser Lys Phe Gly Lys Ser Lys Gly Ile1 5 10 15712PRTArtificial SequenceEngineered peptides based on Ncl protein 157Ile Arg Leu Val Ser Lys Tyr Gly Lys Ser Lys Gly1 5 10 15812PRTArtificial SequenceEngineered peptides based on Ncl protein 158Ile Arg Leu Val Ser Lys Trp Gly Lys Ser Lys Gly1 5 10 15913PRTArtificial SequenceEngineered peptides based on Ncl protein 159Ile Arg Leu Val Ser Lys Leu Trp Gly Lys Ser Lys Gly1 5 10 16032PRTArtificial SequenceEngineered peptides based on Ncl protein 160Lys Gly Gln Asn Gln Xaa Tyr Arg Gly Gly Lys Asn Ser Thr Trp Ser1 5 10 15 Gly Glu Ser Lys Thr Leu Val Leu Ser Asn Leu Ser Tyr Ser Ala Thr 20 25 30 16132PRTArtificial SequenceEngineered peptides based on Ncl protein 161Lys Gly Gln Asn Gln Asp Tyr Arg Gly Gly Lys Asn Ser Thr Trp Ser1 5 10 15 Gly Xaa Ser Lys Thr Leu Val Leu Ser Asn Leu Ser Tyr Ser Ala Thr 20 25 30 16232PRTArtificial SequenceEngineered peptides based on Ncl protein 162Lys Gly Gln Asn Gln Xaa Tyr Arg Gly Gly Lys Asn Ser Thr Trp Ser1 5 10 15 Gly Xaa Ser Lys Thr Leu Val Leu Ser Asn Leu Ser Tyr Ser Ala Thr 20 25 30 16332PRTArtificial SequenceEngineered peptides based on Ncl protein 163Lys Gly Xaa Asn Gln Asp Tyr Arg Leu Gly Lys Asn Ser Thr Trp Ser1 5 10 15 Gly Xaa Ser Lys Thr Leu Val Leu Ser Asn Leu Ser Tyr Ser Ala Thr 20 25 30 16421PRTArtificial SequenceEngineered peptides based on Ncl protein 164Lys Gly Xaa Asn Gln Asp Tyr Arg Leu Gly Lys Asn Ser Thr Trp Ser1 5 10 15 Gly Xaa Ser Lys Thr 20 16521PRTArtificial SequenceEngineered peptides based on Ncl protein 165Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Xaa Lys Ser Lys1 5 10 15 Gly Tyr Ala Phe Ile 20 16621PRTArtificial SequenceEngineered peptides based on Ncl protein 166Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Leu Lys Ser Lys1 5 10 15 Gly Tyr Ala Phe Ile 20 16721PRTArtificial SequenceEngineered peptides based on Ncl protein 167Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Tyr Lys Ser Lys1 5 10 15 Gly Tyr Ala Phe Ile 20 16821PRTArtificial SequenceEngineered peptides based on Ncl protein 168Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Phe Lys Ser Lys1 5 10 15 Gly Tyr Ala Phe Ile 20 16921PRTArtificial SequenceEngineered peptides based on Ncl protein 169Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Trp Lys Ser Lys1 5 10 15 Gly Tyr Ala Phe Ile 20 17018PRTArtificial SequenceEngineered peptides based on Ncl protein 170Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Gly Lys Ser Lys1 5 10 15 Gly Tyr17118PRTArtificial SequenceEngineered peptides based on Ncl protein 171Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Xaa Lys Ser Lys1 5 10 15 Gly Tyr17218PRTArtificial SequenceEngineered peptides based on Ncl protein 172Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Leu Lys Ser Lys1 5 10 15 Gly Tyr17318PRTArtificial SequenceEngineered peptides based on Ncl protein 173Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Tyr Lys Ser Lys1 5 10 15 Gly Tyr17418PRTArtificial SequenceEngineered peptides based on Ncl protein 174Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Phe Lys Ser Lys1 5 10 15 Gly Tyr17518PRTArtificial SequenceEngineered peptides based on Ncl protein 175Lys Ala Thr Phe Ile Lys Val Pro Gln Asn Gln Asn Trp Lys Ser Lys1 5 10 15 Gly Tyr17615PRTArtificial SequenceEngineered peptides based on Csp3 protein 176Ser Lys Ser Ile Lys Asn Leu Glu Pro Lys Ile Ile His Gly Ser1 5 10 15 17731PRTArtificial SequenceEngineered peptides based on Csp3 protein 177Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly Asp Arg Cys Arg Ser Leu1 5 10 15 Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln Ala Cys Arg Gly Thr 20 25 30 17831PRTArtificial SequenceEngineered peptides based on Csp3 protein 178Phe Ile Gln Ser Leu Cys Ala Met Leu Lys Gln Tyr Ala Asp Lys Leu1 5 10 15 Glu Phe Met His Ile Leu Thr Arg Val Asn Arg Lys Val Ala Thr 20 25 30 17915PRTArtificial SequenceEngineered peptides based on Csp3 protein 179Ser Lys Ser Ile Lys Asn Leu Xaa Pro Lys Ile Ile His Gly Ser1 5 10 15 18015PRTArtificial SequenceEngineered peptides based on Csp3 protein 180Ser Lys Ser Ile Lys Asn Leu Glu Pro Lys Ile Ile Tyr Gly Ser1 5 10 15 18115PRTArtificial SequenceEngineered peptides based on Csp3 protein 181Ser Lys Ser Ile Lys Asn Leu Glu Pro Lys Ile Ile Tyr Xaa Ser1 5 10 15 18215PRTArtificial SequenceEngineered peptides based on Csp3 protein 182Ser Lys Ser Ile Lys Asn Leu Xaa Pro Lys Ile Ile Tyr Xaa Ser1 5 10 15 18331PRTArtificial SequenceEngineered peptides based on Csp3 protein 183Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly Xaa Arg Cys Arg Ser Leu1 5 10 15 Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln Ala Cys Arg Gly Thr 20 25 30 18431PRTArtificial SequenceEngineered peptides based on Csp3 protein 184Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly Asp Arg Xaa Arg Ser Leu1 5 10 15 Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln Ala Cys Arg Gly Thr 20 25 30 18531PRTArtificial SequenceEngineered peptides based on Csp3 protein 185Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly Asp Arg Cys Arg Ser Leu1 5 10 15 Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln Ala Xaa Arg Gly Thr 20 25 30 18630PRTArtificial SequenceEngineered peptides based on Csp3 protein 186Leu Lys Lys Ile Thr Asn Phe Arg Gly Xaa Arg Xaa Arg Ser Leu Thr1 5 10 15 Gly Lys Pro Lys Leu Phe Ile Ile Gln Ala Xaa Arg Gly Thr 20 25 30 18718PRTArtificial SequenceEngineered peptides based on Csp3 protein 187Leu Lys Lys Ile Thr Asn Phe Arg Gly Xaa Arg Xaa Arg Ser Leu Thr1 5 10 15 Gly Lys18824PRTArtificial SequenceEngineered peptides based on Bad protein 188Phe Arg Gly Arg Ser Arg Ser Ala Pro Pro Asn Leu Trp Ala Ala Gln1 5 10 15 Arg Tyr Gly Arg Glu Leu Arg Arg 20 18926PRTArtificial SequenceEngineered peptides based on Bad protein 189Arg Arg Met Ser Asp Glu Phe Val Asp Ser Phe Lys Lys Gly Leu Pro1 5 10 15 Arg Pro Lys Ser Ala Gly Thr Ala Thr Gln 20 25 19020PRTArtificial SequenceEngineered peptides based on Bad protein 190Phe Val Asp Ser Phe Lys Lys Gly Leu Pro Arg Pro Lys Ser Ala Gly1 5 10 15 Thr Ala Thr Gln 20 19124PRTArtificial SequenceEngineered peptides based on Bad protein 191Phe Arg Gly Arg Ser Arg Ser Ala Pro Pro Asn Leu Trp Ala Ala Gln1 5 10 15 Arg Tyr Gly Arg Xaa Leu Arg Arg 20 19226PRTArtificial SequenceEngineered peptides based on Bad protein 192Arg Arg Met Ser Xaa Glu Phe Val Asp Ser Phe Lys Lys Gly Leu Pro1 5 10 15 Arg Pro Lys Ser Ala Gly Thr Ala Thr Gln 20 25 19326PRTArtificial SequenceEngineered peptides based on Bad protein 193Arg Arg Met Ser Asp Xaa Phe Val Asp Ser Phe Lys Lys Gly Leu Pro1 5 10 15 Arg Pro Lys Ser Ala Gly Thr Ala Thr Gln 20 25 19426PRTArtificial SequenceEngineered peptides based on Bad protein 194Arg Arg Met Ser Asp Glu Phe Val Xaa Ser Phe Lys Lys Gly Leu Pro1 5 10 15 Arg Pro Lys Ser Ala Gly Thr Ala Thr Gln 20 25 19526PRTArtificial SequenceEngineered peptides based on Bad protein 195Arg Arg Met Ser Xaa Xaa Phe Val Xaa Ser Phe Lys Lys Gly Leu Pro1 5 10 15 Arg Pro Lys Ser Ala Gly Thr Ala Thr Gln 20 25 19626PRTArtificial SequenceEngineered peptides based on Bad protein 196Arg Arg Xaa Ser Xaa Xaa Phe Val Xaa Ser Phe Lys Lys Gly Leu Pro1 5 10 15 Arg Pro Lys Ser Ala Gly Thr Ala Thr Gln 20 25 19720PRTArtificial SequenceEngineered peptides based on Bad protein 197Phe Val Xaa Ser Phe Lys Lys Gly Leu Pro Arg Pro Lys Ser Ala Gly1 5 10 15 Thr Ala Thr Gln 20 19816PRTArtificial SequenceEngineered peptides based on Bad protein 198Phe Val Xaa Ser Phe Lys Lys Gly Leu Xaa Arg Pro Lys Ser Ala Gly1 5 10 15 19916PRTArtificial SequenceEngineered peptides based on Bad protein 199Phe Val Xaa Ser Phe Lys Lys Gly Leu Tyr Arg Pro Lys Ser Ala Gly1 5 10 15 20014PRTArtificial SequenceEngineered peptides based on Prf-1 protein 200Lys Arg Ser His Lys Phe Val Pro Gly Ala Trp Leu Ala Gly1 5 10 20119PRTArtificial SequenceEngineered peptides based on Prf-1 protein 201Val Thr Ser Leu Arg Arg Ser Gly Ser Phe Pro Val Asp Thr Gln Arg1 5 10 15 Phe Leu Arg20216PRTArtificial SequenceEngineered peptides based on Prf-1 protein 202Arg Ser Ile Arg Asn Asp Trp Lys Val Gly Leu Asp Val Thr Pro Lys1 5 10 15 20320PRTArtificial SequenceEngineered peptides based on Prf-1 protein 203Arg Arg Glu Ala Leu Arg Arg Ala Leu Ser Gln Tyr Leu Thr Asp Arg1 5 10 15 Ala Arg Trp Arg 20 20414PRTArtificial SequenceEngineered peptides based on Prf-1 protein 204Asn Leu Asn His Gly His Leu Lys Phe Arg Tyr His Ala Arg1 5 10 20514PRTArtificial SequenceEngineered peptides based on Prf-1 protein 205Lys Arg Ser His Lys Phe Val Xaa Gly Ala Trp Leu Ala Gly1 5 10 20619PRTArtificial SequenceEngineered peptides based on Prf-1 protein 206Val Thr Ser Leu Arg Arg Ser Gly Ser Phe Xaa Val Asp Thr Gln Arg1 5 10 15 Phe Leu Arg20719PRTArtificial SequenceEngineered peptides based on Prf-1 protein 207Val Thr Ser Leu Arg Arg Ser Gly Ser Phe Pro Val Xaa Thr Gln Arg1 5 10 15 Phe Leu Arg20819PRTArtificial SequenceEngineered peptides based on Prf-1 protein 208Val Thr Ser Leu Arg Arg Ser Gly Ser Phe Xaa Val Xaa Thr Gln Arg1 5 10 15 Phe Leu Arg20916PRTArtificial SequenceEngineered peptides based on Prf-1 protein 209Arg Ser Ile Arg Asn Xaa Trp Lys Val Gly Leu Asp Val Thr Pro Lys1 5 10 15 21014PRTArtificial SequenceEngineered peptides based on Prf-1 protein 210Arg Ser Ile Arg Asn Asp Trp Lys Val Gly Leu Asp Val Thr1 5 10 21114PRTArtificial SequenceEngineered peptides based on Prf-1 protein 211Arg Ser Ile Arg Asn Xaa Trp Lys Val Gly Leu Asp Val Thr1 5 10 21220PRTArtificial SequenceEngineered peptides based on Prf-1 protein 212Arg Arg Xaa Ala Leu Arg Arg Ala Leu Ser Gln Tyr Leu Thr Asp Arg1 5 10 15 Ala Arg Trp Arg 20 21320PRTArtificial SequenceEngineered peptides based on Prf-1 protein 213Arg Arg Glu Ala Leu Arg Arg Ala Leu Ser Gln Tyr Leu Thr Xaa Arg1 5 10 15 Ala Arg Trp Arg 20 21420PRTArtificial SequenceEngineered peptides based on Prf-1 protein 214Arg Arg Xaa Ala Leu Arg Arg Ala Leu Ser Gln Tyr Leu Thr Xaa Arg1 5 10 15 Ala Arg Trp Arg 20 21520PRTArtificial SequenceEngineered peptides based on Prf-1 protein 215Arg Xaa Xaa Ala Leu Arg Arg Ala Leu Ser Gln Tyr Leu Thr Xaa Arg1 5 10 15 Ala Arg Trp Arg 20 21614PRTArtificial SequenceEngineered peptides based on Prf-1 protein 216Asn Leu Asn Xaa Gly His Leu Lys Phe Arg Tyr His Ala Arg1 5 10 21714PRTArtificial SequenceEngineered peptides based on Prf-1 protein 217Asn Leu Asn Xaa Gly Xaa Leu Lys Phe Arg Tyr His Ala Arg1 5 10 21814PRTArtificial SequenceEngineered peptides based on Prf-1 protein 218Asn Leu Asn Xaa Gly Xaa Leu Lys Phe Arg Tyr Xaa Ala Arg1 5 10 21914PRTArtificial SequenceEngineered peptides based on Prf-1 protein 219Asn Leu Asn Xaa Gly His Leu Lys Phe Arg Tyr His Ala Arg1 5 10 22014PRTArtificial SequenceEngineered peptides based on Prf-1 protein 220Asn Leu Asn Xaa Gly Xaa Leu Lys Phe Arg Tyr His Ala Arg1 5 10 22114PRTArtificial SequenceEngineered peptides based on Prf-1 protein 221Asn Leu Asn Xaa Gly Xaa Leu Lys Phe Arg Tyr Xaa Ala Arg1 5 10 22214PRTArtificial SequenceEngineered peptides based on Prf-1 protein 222Xaa Leu Asn Xaa Gly Xaa Leu Lys Phe Arg Tyr Xaa Ala Arg1 5 10 22314PRTArtificial SequenceEngineered peptides based on Prf-1 protein 223Xaa Leu Asn Xaa Gly Xaa Leu Lys Phe Arg Tyr Xaa Ala Arg1

5 10 22417PRTArtificial SequenceEngineered peptides based on Granulysin protein 224Leu Gly Arg Asp Tyr Arg Thr Cys Leu Thr Ile Val Gln Lys Leu Lys1 5 10 15 Lys22523PRTArtificial SequenceEngineered peptides based on Granulysin protein 225Lys Pro Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg Val Cys Arg1 5 10 15 Thr Gly Arg Ser Arg Trp Arg 20 22618PRTArtificial SequenceEngineered peptides based on Granulysin protein 226Ser Arg Trp Arg Arg Arg Tyr Gln Ser Arg Val Thr Gln Gly Leu Val1 5 10 15 Ala Gly22717PRTArtificial SequenceEngineered peptides based on Granulysin protein 227Leu Gly Arg Xaa Tyr Arg Thr Cys Leu Thr Ile Val Gln Lys Leu Lys1 5 10 15 Lys22817PRTArtificial SequenceEngineered peptides based on Granulysin protein 228Leu Gly Arg Asp Tyr Arg Thr Xaa Leu Thr Ile Val Gln Lys Leu Lys1 5 10 15 Lys22917PRTArtificial SequenceEngineered peptides based on Granulysin protein 229Leu Gly Arg Xaa Tyr Arg Thr Xaa Leu Thr Ile Val Gln Lys Leu Lys1 5 10 15 Lys23023PRTArtificial SequenceEngineered peptides based on Granulysin protein 230Lys Pro Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg Val Xaa Arg1 5 10 15 Thr Gly Arg Ser Arg Trp Arg 20 23118PRTArtificial SequenceEngineered peptides based on Granulysin protein 231Lys Pro Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg Val Xaa Arg1 5 10 15 Thr Gly23218PRTArtificial SequenceEngineered peptides based on Granulysin protein 232Lys Pro Thr Gln Arg Ser Val Ser Asn Tyr Ala Thr Arg Val Xaa Arg1 5 10 15 Thr Gly23318PRTArtificial SequenceEngineered peptides based on Granulysin protein 233Lys Pro Thr Gln Arg Ser Val Ser Asn Phe Ala Thr Arg Val Xaa Arg1 5 10 15 Thr Gly23417PRTArtificial SequenceEngineered peptides based on Granulysin protein 234Ser Arg Trp Arg Arg Tyr Gln Ser Arg Val Thr Gln Tyr Leu Val Ala1 5 10 15 Gly23517PRTArtificial SequenceEngineered peptides based on Granulysin protein 235Xaa Arg Trp Arg Arg Tyr Gln Ser Arg Val Thr Gln Tyr Leu Val Ala1 5 10 15 Gly23627PRTArtificial SequenceEngineered peptides based on CidA protein 236Gln Lys Ile Phe His Leu Pro Leu Ala Gly Ser Ile Val Gly Leu Phe1 5 10 15 Leu Phe Tyr Leu Leu Leu Gln Phe Lys Ile Val 20 25 23741PRTArtificial SequenceEngineered peptides based on CidA protein 237Glu Ile Thr Leu Asn Tyr Ile Leu Phe Phe Ala Val Ile Ile Ile Gly1 5 10 15 Thr Cys Ile Val Ala Leu Ser Ser Gly Tyr Ile Ala Glu Lys Met Ser 20 25 30 Val Lys His Lys Gln Arg Lys Gly Ile 35 40 23827PRTArtificial SequenceEngineered peptides based on CidA protein 238Gln Lys Ile Phe His Leu Pro Leu Ala Xaa Ser Ile Val Gly Leu Phe1 5 10 15 Leu Phe Tyr Leu Leu Leu Gln Phe Lys Ile Val 20 25 23927PRTArtificial SequenceEngineered peptides based on CidA protein 239Gln Lys Ile Phe His Leu Pro Leu Ala Gly Ser Ile Val Gly Leu Phe1 5 10 15 Leu Phe Tyr Leu Gly Leu Gln Phe Lys Ile Val 20 25 24027PRTArtificial SequenceEngineered peptides based on CidA protein 240Gln Lys Ile Phe His Leu Pro Leu Ala Xaa Ser Ile Val Gly Leu Phe1 5 10 15 Leu Phe Tyr Leu Gly Leu Gln Phe Lys Ile Val 20 25 24120PRTArtificial SequenceEngineered peptides based on CidA protein 241Leu Ala Xaa Ser Ile Val Gly Leu Phe Leu Phe Tyr Leu Gly Leu Gln1 5 10 15 Phe Lys Ile Val 20 24220PRTArtificial SequenceEngineered peptides based on CidA protein 242Leu Ala Xaa Ser Ile Val Xaa Leu Phe Leu Phe Tyr Leu Gly Leu Gln1 5 10 15 Phe Lys Ile Val 20 24341PRTArtificial SequenceEngineered peptides based on CidA protein 243Glu Ile Thr Leu Asn Tyr Ile Leu Phe Phe Ala Val Ile Ile Ile Gly1 5 10 15 Thr Xaa Ile Val Ala Leu Ser Ser Gly Tyr Ile Ala Glu Lys Xaa Ser 20 25 30 Val Lys His Lys Gln Arg Lys Gly Ile 35 40 24414PRTArtificial SequenceEngineered peptides based on CidA protein 244Ala Glu Lys Met Ser Val Lys His Lys Gln Arg Lys Gly Ile1 5 10 24514PRTArtificial SequenceEngineered peptides based on CidA protein 245Ala Xaa Lys Met Ser Val Lys His Lys Gln Arg Lys Gly Ile1 5 10 24614PRTArtificial SequenceEngineered peptides based on CidA protein 246Ala Leu Lys Met Ser Val Lys His Lys Gln Arg Lys Gly Ile1 5 10 24714PRTArtificial SequenceEngineered peptides based on CidA protein 247Ala Leu Lys Xaa Ser Val Lys His Lys Gln Arg Lys Gly Ile1 5 10 24817PRTArtificial SequenceEngineered peptides based on LrgA protein 248Lys Val Thr Ser Arg Ser Lys Gly Asp Lys Val Thr Lys Lys Ile Lys1 5 10 15 Ile24917PRTArtificial SequenceEngineered peptides based on LrgA protein 249Lys Val Thr Ser Arg Ser Lys Gly Asp Lys Val Thr Lys Trp Ile Lys1 5 10 15 Ile25017PRTArtificial SequenceEngineered peptides based on LrgA protein 250Lys Val Thr Ser Arg Ser Lys Gly Asp Lys Val Thr Lys Xaa Ile Lys1 5 10 15 Ile25117PRTArtificial SequenceEngineered peptides based on LrgA protein 251Lys Val Thr Ser Arg Ser Lys Gly Asp Lys Val Thr Lys Xaa Ile Lys1 5 10 15 Ile25228PRTArtificial SequenceEngineered peptides based on Lambda S21 protein 252Ser Leu Val Leu Gly Phe Leu Thr Tyr Leu Thr Asn Leu Tyr Phe Lys1 5 10 15 Ile Arg Glu Asp Arg Arg Lys Ala Ala Arg Gly Glu 20 25 25328PRTArtificial SequenceEngineered peptides based on Lambda S21 protein 253Ser Leu Val Leu Gly Phe Leu Thr Tyr Leu Thr Asn Leu Tyr Phe Lys1 5 10 15 Ile Arg Xaa Asp Arg Arg Lys Ala Ala Arg Gly Glu 20 25 25428PRTArtificial SequenceEngineered peptides based on Lambda S21 protein 254Ser Leu Val Leu Gly Phe Leu Thr Tyr Leu Thr Asn Leu Tyr Phe Lys1 5 10 15 Ile Arg Glu Xaa Arg Arg Lys Ala Ala Arg Gly Glu 20 25 25528PRTArtificial SequenceEngineered peptides based on Lambda S21 protein 255Ser Leu Val Leu Gly Phe Leu Thr Tyr Leu Thr Asn Leu Tyr Phe Lys1 5 10 15 Ile Arg Xaa Xaa Arg Arg Lys Ala Ala Arg Gly Glu 20 25 25615PRTArtificial SequenceEngineered peptides based on Lambda S21 protein 256Leu Tyr Phe Lys Ile Arg Xaa Xaa Arg Arg Lys Ala Ala Arg Gly1 5 10 15 25717PRTArtificial SequenceEngineered peptides based on Holin protein 257Ala Tyr Leu Arg Gly Arg Tyr Asn Gly Gly Ala Phe Thr Lys Thr Val1 5 10 15 Ile25822PRTArtificial SequenceEngineered peptides based on Holin protein 258Ser Ile Gly Ser Leu Ile Lys Arg Phe Ala Ala Lys Lys Ala Gly Val1 5 10 15 Glu Asp Gly Arg Asn Gln 20 25916PRTArtificial SequenceEngineered peptides based on Holin protein 259Ser Ile Gly Ser Leu Ile Lys Arg Phe Ala Ala Lys Lys Ala Gly Val1 5 10 15 26017PRTArtificial SequenceEngineered peptides based on Holin protein 260Xaa Tyr Leu Arg Gly Arg Tyr Asn Gly Gly Ala Phe Thr Lys Thr Val1 5 10 15 Ile26122PRTArtificial SequenceEngineered peptides based on Holin protein 261Ser Ile Gly Ser Leu Ile Lys Arg Phe Ala Ala Lys Lys Ala Gly Val1 5 10 15 Xaa Asp Gly Arg Asn Gln 20 26222PRTArtificial SequenceEngineered peptides based on Holin protein 262Ser Ile Gly Ser Leu Ile Lys Arg Phe Ala Ala Lys Lys Ala Gly Val1 5 10 15 Glu Xaa Gly Arg Asn Gln 20 26316PRTArtificial SequenceEngineered peptides based on Holin protein 263Ser Ile Gly Ser Leu Ile Lys Arg Phe Ala Xaa Lys Lys Ala Gly Val1 5 10 15 26418PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 264Ala Leu Phe Tyr Phe Ala Ser Lys Leu Val Leu Lys Ala Leu Tyr Thr1 5 10 15 Lys Val26514PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 265Ala Leu Lys Tyr Ser Val Lys His Lys Gln Arg Lys Gly Ile1 5 10 26618PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 266Leu Lys Lys Ile Thr Asn Phe Arg Gly Lys Arg Tyr Arg Ser Leu Thr1 5 10 15 Gly Lys26715PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 267Ala Leu Arg Ser Lys Leu Gln Ser Gln Leu Leu Ser Leu Arg Lys1 5 10 15 26816PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 268Ser Ile Gly Ser Leu Ile Lys Arg Phe Ala Tyr Lys Lys Ala Gly Val1 5 10 15 26919PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 269Thr Arg Ala Leu Val Ala Lys Phe Val Gly Tyr Lys Leu Arg Gln Lys1 5 10 15 Gly Tyr Val27026PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 270Asn Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys1 5 10 15 Leu Val Leu Lys Ala Leu Tyr Thr Lys Val 20 25 27119PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 271Thr Val Thr Ile Phe Val Ala Lys Val Leu Thr Ala Ser Leu Thr Ile1 5 10 15 Trp Lys Lys27217PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 272Thr Arg Phe Arg Arg Thr Phe Ser Lys Leu Ala Ala Gln Leu His Val1 5 10 15 Thr27321PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 273Gly Gln Arg Ser Pro Thr Ala Leu Ser Leu Tyr Leu Phe Leu Leu Tyr1 5 10 15 Trp Val Ile Val Lys 20 27423PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 274Lys Lys Ile Glu Val Leu Lys Ser Leu Gln Ser Lys Ala Lys Leu Leu1 5 10 15 Arg Asn Lys Ala Gly Trp Leu 20 27515PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 275Gly Leu Arg Asn Lys Leu Gln Ser Gln Leu Leu Ser Ile Lys Lys1 5 10 15 27620PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 276Lys Leu Ala Lys Lys Val Asp Pro Gln Gly Leu Arg Thr Ile Gly Val1 5 10 15 Ile Thr Lys Leu 20 27715PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 277Lys Ser Lys Lys Tyr Thr Leu Pro Leu Lys Asn Leu Lys Ile Arg1 5 10 15 27815PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 278Ser Lys Ser Ile Lys Asn Leu Lys Pro Lys Ile Ile Tyr Lys Ser1 5 10 15 27920PRTArtificial SequenceNovel therapeutic peptide designs based on programmed cell death effector domains 279Leu Ala Lys Ser Ile Val Arg Leu Phe Leu Phe Tyr Leu Gly Leu Gln1 5 10 15 Phe Lys Ile Val 20 280780PRTHomo sapiensprogrammed cell death / holin-like proteins - Dnm1 (FIG. 1) 280Met Gly Asn Arg Gly Met Glu Asp Leu Ile Pro Leu Val Asn Arg Leu1 5 10 15 Gln Asp Ala Phe Ser Ala Ile Gly Gln Asn Ala Asp Leu Asp Leu Pro 20 25 30 Gln Ile Ala Val Val Gly Gly Gln Ser Ala Gly Lys Ser Ser Val Leu 35 40 45 Glu Asn Phe Val Gly Arg Asp Phe Leu Pro Arg Gly Ser Gly Ile Val 50 55 60 Thr Arg Arg Pro Leu Val Leu Gln Leu Val Asn Ala Thr Thr Glu Tyr65 70 75 80 Ala Glu Phe Leu His Cys Lys Gly Lys Lys Phe Thr Asp Phe Glu Glu 85 90 95 Val Arg Leu Glu Ile Glu Ala Glu Thr Asp Arg Val Thr Gly Thr Asn 100 105 110 Lys Gly Ile Ser Pro Val Pro Ile Asn Leu Arg Val Tyr Ser Pro His 115 120 125 Val Leu Asn Leu Thr Leu Val Asp Leu Pro Gly Met Thr Lys Val Pro 130 135 140 Val Gly Asp Gln Pro Pro Asp Ile Glu Phe Gln Ile Arg Asp Met Leu145 150 155 160 Met Gln Phe Val Thr Lys Glu Asn Cys Leu Ile Leu Ala Val Ser Pro 165 170 175 Ala Asn Ser Asp Leu Ala Asn Ser Asp Ala Leu Lys Val Ala Lys Glu 180 185 190 Val Asp Pro Gln Gly Gln Arg Thr Ile Gly Val Ile Thr Lys Leu Asp 195 200 205 Leu Met Asp Glu Gly Thr Asp Ala Arg Asp Val Leu Glu Asn Lys Leu 210 215 220 Leu Pro Leu Arg Arg Gly Tyr Ile Gly Val Val Asn Arg Ser Gln Lys225 230 235 240 Asp Ile Asp Gly Lys Lys Asp Ile Thr Ala Ala Leu Ala Ala Glu Arg 245 250 255 Lys Phe Phe Leu Ser His Pro Ser Tyr Arg His Leu Ala Asp Arg Met 260 265 270 Gly Thr Pro Tyr Leu Gln Lys Val Leu Asn Gln Gln Leu Thr Asn His 275 280 285 Ile Arg Asp Thr Leu Pro Gly Leu Arg Asn Lys Leu Gln Ser Gln Leu 290 295 300 Leu Ser Ile Glu Lys Glu Val Glu Glu Tyr Lys Asn Phe Arg Pro Asp305 310 315 320 Asp Pro Ala Arg Lys Thr Lys Ala Leu Leu Gln Met Val Gln Gln Phe 325 330 335 Ala Val Asp Phe Glu Lys Arg Ile Glu Gly Ser Gly Asp Gln Ile Asp 340 345 350 Thr Tyr Glu Leu Ser Gly Gly Ala Arg Ile Asn Arg Ile Phe His Glu 355 360 365 Arg Phe Pro Phe Glu Leu Val Lys Met Glu Phe Asp Glu Lys Glu Leu 370 375 380 Arg Arg Glu Ile Ser Tyr Ala Ile Lys Asn Ile His Gly Ile Arg Thr385 390 395 400 Gly Leu Phe Thr Pro Asp Leu Ala Phe Glu Ala Thr Val Lys Lys Gln 405 410 415 Val Gln Lys Leu Lys Glu Pro Ser Ile Lys Cys Val Asp Met Val Val 420 425 430 Ser Glu Leu Thr Ala Thr Ile Arg Lys Cys Ser Glu Lys Leu Gln Gln 435 440 445 Tyr Pro Arg Leu Arg Glu Glu Met Glu Arg Ile Val Thr Thr His Ile 450 455 460 Arg Glu Arg Glu Gly Arg Thr Lys Glu Gln Val Met Leu Leu Ile Asp465 470 475 480 Ile Glu Leu Ala Tyr Met Asn Thr Asn His Glu Asp Phe Ile Gly Phe 485 490 495 Ala Asn Ala Gln Gln Arg Ser Asn Gln Met Asn Lys Lys Lys Thr Ser 500 505 510 Gly Asn Gln Asp Glu Ile Leu Val Ile Arg Lys Gly Trp Leu Thr Ile 515 520 525 Asn Asn Ile Gly Ile Met Lys Gly Gly Ser Lys Glu Tyr Trp Phe Val 530

535 540 Leu Thr Ala Glu Asn Leu Ser Trp Tyr Lys Asp Asp Glu Glu Lys Glu545 550 555 560 Lys Lys Tyr Met Leu Ser Val Asp Asn Leu Lys Leu Arg Asp Val Glu 565 570 575 Lys Gly Phe Met Ser Ser Lys His Ile Phe Ala Leu Phe Asn Thr Glu 580 585 590 Gln Arg Asn Val Tyr Lys Asp Tyr Arg Gln Leu Glu Leu Ala Cys Glu 595 600 605 Thr Gln Glu Glu Val Asp Ser Trp Lys Ala Ser Phe Leu Arg Ala Gly 610 615 620 Val Tyr Pro Glu Arg Val Gly Asp Lys Glu Lys Ala Ser Glu Thr Glu625 630 635 640 Glu Asn Gly Ser Asp Ser Phe Met His Ser Met Asp Pro Gln Leu Glu 645 650 655 Arg Gln Val Glu Thr Ile Arg Asn Leu Val Asp Ser Tyr Met Ala Ile 660 665 670 Val Asn Lys Thr Val Arg Asp Leu Met Pro Lys Thr Ile Met His Leu 675 680 685 Met Ile Asn Asn Thr Lys Glu Phe Ile Phe Ser Glu Leu Leu Ala Asn 690 695 700 Leu Tyr Ser Cys Gly Asp Gln Asn Thr Leu Met Glu Glu Ser Ala Glu705 710 715 720 Gln Ala Gln Arg Arg Asp Glu Met Leu Arg Met Tyr His Ala Leu Lys 725 730 735 Glu Ala Leu Ser Ile Ile Gly Asp Ile Asn Thr Thr Thr Val Ser Thr 740 745 750 Pro Met Pro Pro Pro Val Asp Asp Ser Trp Leu Gln Val Gln Ser Val 755 760 765 Pro Ala Gly Arg Arg Ser Pro Thr Ser Ser Pro Thr 770 775 780 281191PRTHomo sapiensprogrammed cell death / holin-like proteins - Bax (FIG. 1) 281Met Asp Gly Ser Gly Glu Gln Pro Arg Gly Gly Gly Pro Thr Ser Ser1 5 10 15 Glu Gln Ile Met Lys Thr Gly Ala Leu Leu Leu Gln Gly Phe Ile Gln 20 25 30 Asp Arg Ala Gly Arg Met Gly Gly Glu Ala Pro Glu Leu Ala Leu Asp 35 40 45 Pro Val Pro Gln Asp Ala Ser Thr Lys Lys Leu Ser Glu Cys Leu Lys 50 55 60 Arg Ile Gly Asp Glu Leu Asp Ser Asn Met Glu Leu Gln Arg Met Ile65 70 75 80 Ala Ala Val Asp Thr Asp Ser Pro Arg Glu Val Phe Phe Arg Val Ala 85 90 95 Ala Asp Met Phe Ser Asp Gly Asn Phe Asn Trp Gly Arg Val Val Ala 100 105 110 Leu Phe Tyr Phe Ala Ser Lys Val Leu Lys Ala Leu Cys Thr Lys Val 115 120 125 Pro Glu Leu Ile Arg Thr Ile Met Gly Trp Thr Leu Asp Phe Leu Arg 130 135 140 Glu Arg Leu Leu Gly Trp Ile Gln Asp Gln Gly Gly Trp Asp Gly Leu145 150 155 160 Leu Ser Tyr Phe Gly Thr Pro Thr Trp Gln Thr Val Thr Ile Phe Val 165 170 175 Ala Gly Val Leu Thr Ala Ser Leu Thr Ile Trp Lys Lys Met Gly 180 185 190 282166PRTHomo sapiensprogrammed cell death / holin-like proteins - Bcl-2 (FIG. 1) 282Met Ala His Ala Gly Arg Thr Gly Tyr Asp Asn Arg Glu Ile Val Met1 5 10 15 Lys Tyr Ile His Tyr Lys Leu Ser Gln Arg Gly Tyr Glu Trp Asp Ala 20 25 30 Gly Asp Asp Val Glu Glu Asn Arg Thr Glu Ala Pro Glu Gly Thr Glu 35 40 45 Ser Glu Val Val His Leu Ala Leu Arg Gln Ala Gly Asp Asp Phe Ser 50 55 60 Arg Arg Tyr Arg Gly Asp Phe Ala Glu Met Ser Ser Gln Leu His Leu65 70 75 80 Thr Pro Phe Thr Ala Arg Gly Arg Phe Ala Thr Val Val Glu Glu Leu 85 90 95 Phe Arg Asp Gly Val Asn Trp Gly Arg Ile Val Ala Phe Phe Glu Phe 100 105 110 Gly Gly Val Met Cys Val Glu Ser Val Asn Arg Glu Met Ser Pro Leu 115 120 125 Val Asp Asn Ile Ala Leu Trp Met Thr Glu Tyr Leu Asn Arg His Leu 130 135 140 His Thr Trp Ile Gln Asp Asn Gly Gly Trp Asp Ala Phe Val Glu Leu145 150 155 160 Tyr Gly Pro Ser Met Xaa 165 283131PRTStaphylococcus aureusprogrammed cell death / holin-like proteins - CidA (FIG. 2) 283Met His Lys Val Gln Leu Ile Ile Lys Leu Leu Leu Gln Leu Gly Ile1 5 10 15 Ile Ile Val Ile Thr Tyr Ile Gly Thr Glu Ile Gln Lys Ile Phe His 20 25 30 Leu Pro Leu Ala Gly Ser Ile Val Gly Leu Phe Leu Phe Tyr Leu Leu 35 40 45 Leu Gln Phe Lys Ile Val Pro Leu Thr Trp Val Glu Asp Gly Ala Asn 50 55 60 Phe Leu Leu Lys Thr Met Val Phe Phe Phe Ile Pro Ser Val Val Gly65 70 75 80 Ile Met Asp Val Ala Ser Glu Ile Thr Leu Asn Tyr Ile Leu Phe Phe 85 90 95 Ala Val Ile Ile Ile Gly Thr Cys Ile Val Ala Leu Ser Ser Gly Tyr 100 105 110 Ile Ala Glu Lys Met Ser Val Lys His Lys Gln Arg Lys Gly Ile Asp 115 120 125 Ala Tyr Glu 130 284147PRTStaphylococcus aureusprogrammed cell death proteins - LrgA (FIG. 2) 284Met Val Val Lys Gln Gln Lys Asp Ala Ser Lys Pro Ala His Phe Phe1 5 10 15 His Gln Val Ile Val Ile Ala Leu Val Leu Phe Val Ser Lys Ile Ile 20 25 30 Glu Ser Phe Met Pro Ile Pro Met Pro Ala Ser Val Ile Gly Leu Val 35 40 45 Leu Leu Phe Val Leu Leu Cys Thr Gly Ala Val Lys Leu Gly Glu Val 50 55 60 Glu Lys Val Gly Thr Thr Leu Thr Asn Asn Ile Gly Leu Leu Phe Val65 70 75 80 Pro Ala Gly Ile Ser Val Val Asn Ser Leu Gly Val Ile Ser Gln Ala 85 90 95 Pro Phe Leu Ile Ile Gly Leu Ile Ile Val Ser Thr Ile Leu Leu Leu 100 105 110 Ile Cys Thr Gly Tyr Val Thr Gln Ile Ile Met Lys Val Thr Ser Arg 115 120 125 Ser Lys Gly Asp Lys Val Thr Lys Lys Ile Lys Ile Glu Glu Ala Gln 130 135 140 Ala His Asp145 285309PRTBos taurusPerforin 1 protein (Fig. 3) 285Asp Thr Gln Arg Phe Leu Arg Pro Asp Gly Thr Cys Thr Leu Cys Arg1 5 10 15 Asn Ala Leu Gln Lys Asp Val Leu Gln Arg Leu Pro Leu Ala Ile Thr 20 25 30 Asp Trp Arg Ala His Gly Ala Gly Cys Lys Arg Arg Val Val Lys Leu 35 40 45 Glu Gly Arg Ser Thr Glu Asp Val Ala Gly Glu Ala Ala Asn Arg Ile 50 55 60 Arg Asn Asp Trp Gln Val Gly Leu Asp Val Ser Pro Lys Pro Asn Ala65 70 75 80 Asn Val Arg Val Thr Val Ala Gly Ser His Ser Glu Asp Ala Asn Phe 85 90 95 Ala Ala Gln Lys Thr His Gln Asp Asn Tyr Arg Phe Ser Met Asp Leu 100 105 110 Val Glu Cys Arg Phe Tyr Ser Phe His Leu Val His Thr Pro Pro Val 115 120 125 His Pro Glu Phe Lys Arg Ala Leu Lys Thr Leu Pro Pro His Phe Asn 130 135 140 Thr Ser Thr Lys Pro Asp Tyr His Arg Leu Ile Ser Ser Tyr Gly Thr145 150 155 160 His Phe Ile Arg Ser Met Glu Leu Gly Gly Arg Ile Ser Ala Leu Thr 165 170 175 Ala Leu Arg Thr Cys Glu Leu Ala Leu Glu Gly Leu Thr Ala Ser Glu 180 185 190 Val Glu Asp Cys Leu Ala Val Glu Ala Glu Val Ser Ile Ser Asp Arg 195 200 205 Ala Ser Ala Ser Pro Ser Phe Lys Ala Cys Glu Glu Lys Lys Lys Asn 210 215 220 His Lys Val Gly Thr Ser Phe His Gln Ala Tyr Arg Glu Arg His Ser225 230 235 240 Asn Val Asp Gly Gly His His Ser Thr Met His Asp Leu Leu Phe Gly 245 250 255 Ser Gln Ala Gly Pro Glu Gln Phe Ser Ala Trp Val Ala Ser Leu Gln 260 265 270 Asp Ser Pro Gly Leu Val Asp Tyr Thr Leu Glu Pro Leu His Met Leu 275 280 285 Val Glu Ser Gln Asp Pro Arg Arg Glu Ala Leu Arg Gln Ala Val Ser 290 295 300 Lys Tyr Val Thr Asp305 286239PRTHomo sapiensBcl-2 protein (Fig. 3) 286Met Ala His Ala Gly Arg Thr Gly Tyr Asp Asn Arg Glu Ile Val Met1 5 10 15 Lys Tyr Ile His Tyr Lys Leu Ser Gln Arg Gly Tyr Glu Trp Asp Ala 20 25 30 Gly Asp Val Gly Ala Ala Pro Pro Gly Ala Ala Pro Ala Pro Gly Ile 35 40 45 Phe Ser Ser Gln Pro Gly His Thr Pro His Pro Ala Ala Ser Arg Asp 50 55 60 Pro Val Ala Arg Thr Ser Pro Leu Gln Thr Pro Ala Ala Pro Gly Ala65 70 75 80 Ala Ala Gly Pro Ala Leu Ser Pro Val Pro Pro Val Val His Leu Thr 85 90 95 Leu Arg Gln Ala Gly Asp Asp Phe Ser Arg Arg Tyr Arg Arg Asp Phe 100 105 110 Ala Glu Met Ser Ser Gln Leu His Leu Thr Pro Phe Thr Ala Arg Gly 115 120 125 Arg Phe Ala Thr Val Val Glu Glu Leu Phe Arg Asp Gly Val Asn Trp 130 135 140 Gly Arg Ile Val Ala Phe Phe Glu Phe Gly Gly Val Met Cys Val Glu145 150 155 160 Ser Val Asn Arg Glu Met Ser Pro Leu Val Asp Asn Ile Ala Leu Trp 165 170 175 Met Thr Glu Tyr Leu Asn Arg His Leu His Thr Trp Ile Gln Asp Asn 180 185 190 Gly Gly Trp Asp Ala Phe Val Glu Leu Tyr Gly Pro Ser Met Arg Pro 195 200 205 Leu Phe Asp Phe Ser Trp Leu Ser Leu Lys Thr Leu Leu Ser Leu Ala 210 215 220 Leu Val Gly Ala Cys Ile Thr Leu Gly Ala Tyr Leu Gly His Lys225 230 235 287193PRTHomo sapiensBCL-W protein (Fig. 3) 287Met Ala Thr Pro Ala Ser Ala Pro Asp Thr Arg Ala Leu Val Ala Asp1 5 10 15 Phe Val Gly Tyr Lys Leu Arg Gln Lys Gly Tyr Val Cys Gly Ala Gly 20 25 30 Pro Gly Glu Gly Pro Ala Ala Asp Pro Leu His Gln Ala Met Arg Ala 35 40 45 Ala Gly Asp Glu Phe Glu Thr Arg Phe Arg Arg Thr Phe Ser Asp Leu 50 55 60 Ala Ala Gln Leu His Val Thr Pro Gly Ser Ala Gln Gln Arg Phe Thr65 70 75 80 Gln Val Ser Asp Glu Leu Phe Gln Gly Gly Pro Asn Trp Gly Arg Leu 85 90 95 Val Ala Phe Phe Val Phe Gly Ala Ala Leu Cys Ala Glu Ser Val Asn 100 105 110 Lys Glu Met Glu Pro Leu Val Gly Gln Val Gln Glu Trp Met Val Ala 115 120 125 Tyr Leu Glu Thr Arg Leu Ala Asp Trp Ile His Ser Ser Gly Gly Trp 130 135 140 Ala Glu Phe Thr Ala Leu Tyr Gly Asp Gly Ala Leu Glu Glu Ala Arg145 150 155 160 Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val Leu Thr Gly 165 170 175 Ala Val Ala Leu Gly Ala Leu Val Thr Val Gly Ala Phe Phe Ala Ser 180 185 190 Lys 28819PRTArtificial Sequenceengineered peptide based on programmed cell death effector proteins 288Ser Gln Ser Asn Arg Glu Leu Val Val Asp Phe Leu Ser Tyr Lys Leu1 5 10 15 Ser Gln Lys28917PRTArtificial Sequenceengineered peptide based on programmed cell death effector proteins 289Gln Lys Leu Lys Lys Met Val Asp Lys Pro Thr Gln Arg Ser Val Ser1 5 10 15 Asn29076PRTHomo sapiensHelix 1 of human Bcl-2 290Met Ala His Ala Gly Arg Thr Gly Tyr Asp Asn Arg Glu Ile Val Met1 5 10 15 Lys Tyr Ile His Tyr Lys Leu Ser Gln Arg Gly Tyr Glu Trp Asp Ala 20 25 30 Gly Asp Asp Val Glu Glu Asn Arg Thr Glu Ala Pro Glu Gly Thr Glu 35 40 45 Ser Glu Val Val His Leu Thr Leu Arg Gln Ala Gly Asp Asp Phe Ser 50 55 60 Arg Arg Tyr Arg Arg Asp Phe Ala Glu Met Ser Ser65 70 75 29176PRTHomo sapiensHelix 2 of human Bcl-2 291Gly Asp Asp Val Glu Glu Asn Arg Thr Glu Ala Pro Glu Gly Thr Glu1 5 10 15 Ser Glu Val Val His Leu Ala Leu Arg Gln Ala Gly Asp Asp Phe Ser 20 25 30 Arg Arg Tyr Arg Gly Asp Phe Ala Glu Met Ser Ser Gln Leu His Leu 35 40 45 Thr Pro Phe Thr Ala Arg Gly Arg Phe Ala Thr Val Val Glu Glu Leu 50 55 60 Phe Arg Asp Gly Val Asn Trp Gly Arg Ile Val Ala65 70 75 29277PRTHomo sapiensHelix 1 of human Bcl-X1 292Met Ser Gln Ser Asn Arg Glu Leu Val Val Asp Phe Leu Ser Tyr Lys1 5 10 15 Leu Ser Gln Lys Gly Tyr Ser Trp Ser Gln Phe Ser Asp Val Glu Glu 20 25 30 Asn Arg Thr Glu Ala Pro Glu Gly Thr Glu Ser Glu Met Glu Thr Pro 35 40 45 Ser Ala Ile Asn Gly Asn Pro Ser Trp His Lys Leu Ala Asp Ser Pro 50 55 60 Ala Val Asn Gly Ala Thr Gly His Ser Ser Ser Leu Asp65 70 75 29377PRTHomo sapiensHelix 2 of human Bcl-W 293Gly Pro Leu Gly Ser Met Ala Thr Pro Ala Ser Ala Pro Asp Thr Arg1 5 10 15 Ala Leu Val Ala Asp Phe Val Gly Tyr Lys Leu Arg Gln Lys Gly Tyr 20 25 30 Val Cys Gly Ala Gly Pro Gly Glu Gly Pro Ala Ala Asp Pro Leu His 35 40 45 Gln Ala Met Arg Ala Ala Gly Asp Glu Phe Glu Thr Arg Phe Arg Arg 50 55 60 Thr Phe Ser Asp Leu Ala Ala Gln Leu His Val Thr Pro65 70 75 29474PRTHomo sapiensHelix 4 of human Bax 294Pro Arg Glu Val Phe Phe Arg Val Ala Ala Asp Met Phe Ser Asp Gly1 5 10 15 Asn Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys 20 25 30 Leu Val Leu Lys Ala Leu Cys Thr Lys Val Pro Glu Leu Ile Arg Thr 35 40 45 Ile Met Gly Trp Thr Leu Asp Phe Leu Arg Glu Arg Leu Leu Gly Trp 50 55 60 Ile Gln Asp Gln Gly Gly Trp Asp Gly Leu65 70 29574PRTHomo sapiensHelix 1 of human CTL Granulysin 295Gly Arg Asp Tyr Arg Thr Cys Leu Thr Ile Val Gln Lys Leu Lys Lys1 5 10 15 Met Val Asp Lys Pro Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg 20 25 30 Val Cys Arg Thr Gly Arg Ser Arg Trp Arg Asp Val Cys Arg Asn Phe 35 40 45 Met Arg Arg Tyr Gln Ser Arg Val Ile Gln Gly Leu Val Ala Gly Glu 50 55 60 Thr Ala Gln Gln Ile Cys Glu Asp Leu Arg65 70 29674PRTHomo sapiensHelix 1-2 span of human CTL Granulysin 296Gly Arg Asp Tyr Arg Thr Cys Leu Thr Ile Val Gln Lys Leu Lys Lys1 5 10 15 Met Val Asp Lys Pro Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg 20 25 30 Val Cys Arg Thr Gly Arg Ser Arg Trp Arg Asp Val Cys Arg Asn Phe 35 40 45 Met Arg Arg Tyr Gln Ser Arg Val Ile Gln Gly Leu Val Ala Gly Glu 50 55 60 Thr Ala Gln Gln Ile Cys Glu Asp Leu Arg65

70 29718PRTHomo sapiens 297Phe Thr Ala Arg Gly Arg Phe Ala Thr Val Val Glu Glu Leu Phe Arg1 5 10 15 Asp Gly29818PRTHomo sapiens 298Gly Ser Ala Gln Gln Arg Phe Thr Gln Val Ser Asp Glu Leu Phe Gln1 5 10 15 Gly Gly29918PRTHomo sapiens 299Ser Pro Arg Glu Val Phe Phe Arg Val Ala Ala Asp Met Phe Ser Asp1 5 10 15 Gly Asn30050PRTHomo sapiens 300Glu Glu Val Tyr Lys Arg Pro Leu Phe Leu Gln Pro Thr Tyr Arg Tyr1 5 10 15 His Arg Leu Pro Leu Pro Glu Gln Gly Ser Pro Leu Glu Ala Gln Leu 20 25 30 Asp Ala Phe Val Ser Val Leu Arg Glu Thr Pro Ser Leu Leu Gln Leu 35 40 45 Arg Asp 50 30119PRTHomo sapiens 301Glu Gly Pro Gln Gly Asp Leu Leu Thr Lys Thr Gln Glu Leu Gly Arg1 5 10 15 Asp Tyr Arg30226PRTHomo sapiens 302Glu Val Arg Ala Gln Leu Leu Glu Leu Pro Tyr Ala Arg Lys Glu Leu1 5 10 15 Ser Leu Leu Val Leu Leu Pro Asp Asp Gly 20 25 30320PRTHomo sapiens 303Leu Ile Ser Ser Tyr Gly Thr His Phe Ile Arg Ser Met Glu Leu Gly1 5 10 15 Gly Arg Ile Ser 20 30420PRTHomo sapiens 304Leu Gln Asn Ala Thr Val Glu Ala Gly Thr Arg Cys Gln Val Ala Gly1 5 10 15 Trp Gly Ser Gln 20 30520PRTHomo sapiens 305Ser Ser Lys Ala Gln Val Lys Pro Gly Gln Leu Cys Ser Val Ala Gly1 5 10 15 Trp Gly Tyr Val 20 30620PRTHomo sapiens 306Ser Ala Pro His Gln Pro Gly Pro Ser Leu Trp Ala Glu Ala Lys Thr1 5 10 15 Ser Glu Ala Pro 20 30721PRTHomo sapiens 307Lys Asp Gly Val Thr Pro Ile Lys Asp Leu Thr Ala His Phe Arg Gly1 5 10 15 Asp Arg Cys Lys Thr 20 30819PRTHomo sapiens 308Pro Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His Leu Thr Gly1 5 10 15 Lys Ser Asn30928PRTHomo sapiens 309Val Asn Trp Gly Arg Ile Val Ala Phe Phe Glu Phe Gly Gly Val Met1 5 10 15 Cys Val Glu Ser Val Asn Arg Glu Met Ser Pro Leu 20 25 31028PRTHomo sapiens 310Pro Asn Trp Gly Arg Leu Val Ala Phe Phe Val Phe Gly Ala Ala Leu1 5 10 15 Cys Ala Glu Ser Val Asn Lys Glu Met Glu Pro Leu 20 25 31128PRTHomo sapiens 311Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu1 5 10 15 Val Leu Lys Ala Leu Cys Thr Lys Val Pro Glu Leu 20 25 31250PRTHomo sapiens 312Ala His Gly Pro Pro Pro Ala Leu Val Phe Ser Cys Gln Met Gly Val1 5 10 15 Gly Arg Thr Asn Leu Gly Met Val Leu Gly Thr Leu Ile Leu Leu His 20 25 30 Arg Ser Gly Thr Thr Ser Gln Pro Glu Ala Ala Pro Thr Gln Ala Lys 35 40 45 Pro Leu 50 31328PRTHomo sapiens 313Thr Cys Leu Thr Ile Val Gln Lys Leu Lys Lys Met Val Asp Lys Pro1 5 10 15 Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg Val 20 25 31431PRTHomo sapiens 314Val Glu Leu Ser Thr Val Glu Lys Ser Leu Thr Phe Glu Lys Leu Thr1 5 10 15 Ala Trp Thr Lys Pro Asp Cys Met Lys Ser Thr Glu Val Glu Val 20 25 30 31531PRTHomo sapiens 315Ala Leu Thr Ala Leu Arg Thr Cys Glu Leu Ala Leu Glu Gly Leu Thr1 5 10 15 Ala Ser Glu Val Glu Asp Cys Leu Ala Val Glu Ala Glu Val Ser 20 25 30 31628PRTHomo sapiens 316Arg Ser Gly Gly Arg Leu Ser Arg Phe Pro Arg Phe Val Asn Val Thr1 5 10 15 Val Thr Pro Glu Asp Gln Cys Arg Pro Asn Asn Val 20 25 31725PRTHomo sapiens 317Ser Met Ser Thr Leu Ala Thr Thr Leu Gln Glu Val Leu Leu Thr Val1 5 10 15 Gln Lys Asp Cys Gln Cys Glu Arg Leu 20 25 31828PRTHomo sapiens 318Ser Thr Gln Asp Pro Ser Thr Gln Ala Ser Thr Ala Ser Ser Pro Ala1 5 10 15 Pro Glu Glu Asn Ala Pro Ser Glu Gly Gln Arg Val 20 25 31929PRTHomo sapiens 319Leu Leu Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Arg Gly Thr1 5 10 15 Glu Leu Asp Asp Gly Ile Gln Ala Asp Ser Gly Pro Ile 20 25 32027PRTHomo sapiens 320Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu1 5 10 15 Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 20 25 32115PRTHomo sapiens 321Val Asp Asn Ile Ala Leu Trp Met Thr Glu Tyr Leu Asn Arg His1 5 10 15 32215PRTHomo sapiens 322Val Gly Gln Val Gln Glu Trp Met Val Ala Tyr Leu Glu Thr Arg1 5 10 15 32315PRTHomo sapiens 323Ile Arg Thr Ile Met Gly Trp Thr Leu Asp Phe Leu Arg Glu Arg1 5 10 15 32450PRTHomo sapiens 324Pro Met Glu Gln Phe Gln Val Ile Gln Ser Phe Leu Arg Met Val Pro1 5 10 15 Gln Gly Arg Arg Met Val Glu Glu Val Asp Arg Ala Ile Thr Ala Cys 20 25 30 Ala Glu Leu His Asp Leu Lys Glu Val Val Leu Glu Asn Gln Lys Lys 35 40 45 Leu Glu 50 32515PRTHomo sapiens 325Cys Arg Thr Gly Arg Ser Arg Trp Arg Asp Val Cys Arg Asn Phe1 5 10 15 32621PRTHomo sapiens 326Leu Leu Pro Lys Phe Lys Leu Gln Glu Asp Tyr Asp Met Glu Ser Val1 5 10 15 Leu Arg His Leu Gly 20 32724PRTHomo sapiens 327Ile Ser Asp Arg Ala Ser Ala Ser Pro Ser Phe Lys Ala Cys Glu Glu1 5 10 15 Lys Lys Lys Asn His Lys Val Gly 20 32815PRTHomo sapiens 328Cys Thr Gly Val Leu Thr Arg Arg Gly Gly Ile Cys Asn Gly Asp1 5 10 15 32924PRTHomo sapiens 329Phe His Gly Asn Tyr Ser Arg Ala Thr Glu Ile Cys Val Gly Asp Pro1 5 10 15 Lys Lys Thr Gln Thr Gly Phe Lys 20 33025PRTHomo sapiens 330Trp Gly Gln Gly Gln Ser Pro Arg Pro Glu Asn Ser Leu Glu Arg Glu1 5 10 15 Glu Met Gly Pro Val Pro Ala His Thr 20 25 33122PRTHomo sapiens 331Asn Asp Thr Asp Ala Asn Pro Arg Tyr Lys Ile Pro Val Glu Ala Asp1 5 10 15 Phe Leu Phe Ala Tyr Ser 20 33225PRTHomo sapiens 332Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe1 5 10 15 Arg Gly Gln Ser Cys Asn Asn Leu Pro 20 25 33353PRTHomo sapiens 333Pro Leu Gln Thr Pro Ala Ala Pro Gly Ala Ala Ala Gly Pro Ala Leu1 5 10 15 Ser Pro Val Pro Pro Val Val His Leu Thr Leu Arg Gln Ala Gly Asp 20 25 30 Asp Phe Ser Arg Arg Tyr Arg Arg Asp Phe Ala Glu Met Ser Ser Gln 35 40 45 Leu His Leu Thr Pro 50 33434PRTHomo sapiens 334Ala Asp Pro Leu His Gln Ala Met Arg Ala Ala Gly Asp Glu Phe Glu1 5 10 15 Thr Arg Phe Arg Arg Thr Phe Ser Asp Leu Ala Ala Gln Leu His Val 20 25 30 Thr Pro33538PRTHomo sapiens 335Pro Val Pro Gln Asp Ala Ser Thr Lys Lys Leu Ser Glu Cys Leu Lys1 5 10 15 Arg Ile Gly Asp Glu Leu Asp Ser Asn Met Glu Leu Gln Arg Met Ile 20 25 30 Ala Ala Val Asp Thr Asp 35 33682PRTHomo sapiens 336Glu Pro Val Leu Phe Leu Arg Ala Asp Glu Asp Phe Val Ser Tyr Thr1 5 10 15 Pro Arg Asp Lys Gln Asn Leu His Glu Asn Leu Gln Gly Leu Gly Pro 20 25 30 Gly Val Arg Val Glu Ser Leu Glu Leu Ala Ile Arg Lys Glu Ile His 35 40 45 Asp Phe Ala Gln Leu Ser Glu Asn Thr Tyr His Val Tyr His Asn Thr 50 55 60 Glu Asp Leu Trp Gly Glu Pro His Ala Val Ala Ile His Gly Glu Asp65 70 75 80 Asp Leu33729PRTHomo sapiens 337Leu Val Phe Ser Arg Leu Ser Pro Glu Tyr Tyr Asp Pro Ala Arg Ala1 5 10 15 His Leu Arg Asp Gly Glu Lys Ser Cys Pro Cys Gly Gln 20 25 33861PRTHomo sapiens 338Gly His Glu Ala Phe Leu Leu Thr Glu Gly Ser Glu Glu Lys Arg Ser1 5 10 15 Ala Lys Thr Val Asn Gln Leu Ala His Ala Leu His Gln Asp Lys Gln 20 25 30 Leu His Ala Gly Ser Leu Val Ser Val Met Trp Pro Asn Ser Lys Cys 35 40 45 Pro Leu Leu Lys Asp Asp Leu Val Leu Met Asp Ser Pro 50 55 60 33947PRTHomo sapiens 339Pro Val Ser Ile Ser Ser Ala Leu Ala Met Val Leu Leu Gly Ala Lys1 5 10 15 Gly Asn Thr Ala Thr Gln Met Ala Gln Ala Leu Ser Leu Asn Thr Glu 20 25 30 Glu Asp Ile His Arg Ala Phe Gln Ser Leu Leu Thr Glu Val Asn 35 40 45 34077PRTHomo sapiens 340Ile Leu Ala Val Ser Pro Ala Asn Ser Asp Leu Ala Asn Ser Asp Ala1 5 10 15 Leu Lys Val Ala Lys Glu Val Asp Pro Gln Gly Gln Arg Thr Ile Gly 20 25 30 Val Ile Thr Lys Leu Asp Leu Met Asp Glu Gly Thr Asp Ala Arg Asp 35 40 45 Val Leu Glu Asn Lys Leu Leu Pro Leu Arg Arg Gly Tyr Ile Gly Val 50 55 60 Val Asn Arg Ser Gln Lys Asp Ile Asp Gly Lys Lys Asp65 70 75 34185PRTHomo sapiens 341Pro Ser Leu Pro Ser Gln Ala Val Trp Ser Gln Gly Pro Pro Pro Pro1 5 10 15 Pro Pro Tyr Gly Arg Leu Leu Ala Asn Ser Asn Ala His Pro Gly Pro 20 25 30 Phe Pro Pro Ser Thr Gly Ala Gln Ser Thr Ala His Pro Pro Val Ser 35 40 45 Thr His His His His His Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 50 55 60 Gln Gln Gln Gln Gln Gln Gln Gln Gln His His Gly Asn Ser Gly Pro65 70 75 80 Pro Pro Pro Gly Ala 85 34252PRTHomo sapiens 342Leu Cys Leu Leu Val Met Phe Phe Met Val Leu Val Ala Leu Val Gly1 5 10 15 Leu Gly Leu Gly Met Phe Gln Leu Phe His Leu Gln Lys Glu Leu Ala 20 25 30 Glu Leu Arg Glu Ser Thr Ser Gln Met His Thr Ala Ser Ser Leu Glu 35 40 45 Lys Gln Ile Gly 50 34358PRTHomo sapiens 343Phe Ala Ala Gln Lys Thr His Gln Asp Asn Tyr Arg Phe Ser Met Asp1 5 10 15 Leu Val Glu Cys Arg Phe Tyr Ser Phe His Leu Val His Thr Pro Pro 20 25 30 Val His Pro Glu Phe Lys Arg Ala Leu Lys Thr Leu Pro Pro His Phe 35 40 45 Asn Thr Ser Thr Lys Pro Asp Tyr His Arg 50 55 34458PRTHomo sapiens 344Met Asp Glu Ser Val Val Leu Glu Pro Glu Ala Thr Gly Glu Ser Ser1 5 10 15 Ser Leu Glu Pro Thr Pro Ser Ser Gln Glu Ala Gln Arg Ala Leu Gly 20 25 30 Thr Ser Pro Glu Leu Pro Thr Gly Val Thr Gly Ser Ser Gly Thr Arg 35 40 45 Leu Pro Pro Thr Pro Lys Ala Gln Asp Gly 50 55 34544PRTHomo sapiens 345Ala Tyr Asp Leu Arg Arg Arg Glu Arg Gln Ser Arg Gln Thr Phe Ser1 5 10 15 Ile Ser Ser Met Ser Glu Asn Gly Tyr Asp Pro Gln Gln Asn Leu Asn 20 25 30 Asp Leu Met Leu Leu Gln Leu Asp Arg Glu Ala Asn 35 40 34644PRTHomo sapiens 346Ala His Asn Ile Lys Glu Gln Glu Arg Thr Gln Gln Phe Ile Pro Val1 5 10 15 Lys Arg Pro Ile Pro His Pro Ala Tyr Asn Pro Lys Asn Phe Ser Asn 20 25 30 Asp Ile Met Leu Leu Gln Leu Glu Arg Lys Ala Lys 35 40 34763PRTHomo sapiens 347Glu Asp Asp Glu Glu Glu Asp Glu Phe Glu Pro Pro Ile Val Lys Gly1 5 10 15 Val Lys Pro Ala Lys Ala Ala Pro Ala Ala Pro Ala Ser Glu Asp Glu 20 25 30 Glu Asp Asp Glu Asp Glu Asp Asp Glu Glu Asp Asp Asp Glu Glu Glu 35 40 45 Glu Asp Asp Ser Glu Glu Glu Val Met Glu Ile Thr Thr Ala Lys 50 55 60 34847PRTHomo sapiens 348Phe Asp Val Ile Val Tyr Asn Asp Cys Ser Cys Ala Lys Met Gln Asp1 5 10 15 Leu Leu Lys Lys Ala Ser Glu Glu Asp His Thr Asn Ala Ala Cys Phe 20 25 30 Ala Cys Ile Leu Leu Ser His Gly Glu Glu Asn Val Ile Tyr Gly 35 40 45 34918PRTHomo sapiens 349Phe Thr Ala Arg Gly Arg Phe Ala Thr Val Val Glu Glu Leu Phe Arg1 5 10 15 Asp Gly35018PRTHomo sapiens 350Gly Ser Ala Gln Gln Arg Phe Thr Gln Val Ser Asp Glu Leu Phe Gln1 5 10 15 Gly Gly35118PRTHomo sapiens 351Ser Pro Arg Glu Val Phe Phe Arg Val Ala Ala Asp Met Phe Ser Asp1 5 10 15 Gly Asn35270PRTHomo sapiens 352Val Ala Ile His Gly Glu Asp Asp Leu His Val Thr Glu Glu Val Tyr1 5 10 15 Lys Arg Pro Leu Phe Leu Gln Pro Thr Tyr Arg Tyr His Arg Leu Pro 20 25 30 Leu Pro Glu Gln Gly Ser Pro Leu Glu Ala Gln Leu Asp Ala Phe Val 35 40 45 Ser Val Leu Arg Glu Thr Pro Ser Leu Leu Gln Leu Arg Asp Ala His 50 55 60 Gly Pro Pro Pro Ala Leu65 70 35319PRTHomo sapiens 353Glu Gly Pro Gln Gly Asp Leu Leu Thr Lys Thr Gln Glu Leu Gly Arg1 5 10 15 Asp Tyr Arg35444PRTHomo sapiens 354Gly Ile Asp Val Thr Thr Glu Leu Asp Ser Trp Ile Asp Lys Phe Cys1 5 10 15 Leu Asp Ala Asp Val Phe Val Leu Val Ala Asn Ser Glu Ser Thr Leu 20 25 30 Met Gln Thr Glu Lys His Phe Phe His Lys Val Ser 35 40 35519PRTHomo sapiens 355Lys Ala Gly Thr Gln Tyr Leu Leu Arg Thr Ala Asn Arg Leu Phe Gly1 5 10 15 Glu Lys Thr35657PRTHomo sapiens 356Gln Lys Asp Ile Asp Gly Lys Lys Asp Ile Thr Ala Ala Leu Ala Ala1 5 10 15 Glu Arg Lys Phe Phe Leu Ser His Pro Ser Tyr Arg His Leu Ala Asp 20 25 30 Arg Met Gly Thr Pro Tyr Leu Gln Lys Val Leu Asn Gln Gln Leu Thr 35 40 45 Asn His Ile Arg Asp Thr Leu Pro Gly 50 55 35770PRTHomo sapiens 357Asn Ser Gly Pro Pro Pro Pro Gly Ala Phe Pro His Pro Leu Glu Gly1 5 10 15 Gly Ser Ser His His Ala His Pro Tyr Ala Met Ser Pro Ser Leu Gly 20 25 30 Ser Leu Arg Pro Tyr Pro Pro Gly Pro Ala His Leu Pro Pro Pro His 35 40 45 Ser Gln Val Ser Tyr Ser Gln Ala Gly Pro Asn Gly Pro Pro Val Ser 50 55 60 Ser Ser Ser Asn Ser Ser65 70 35822PRTHomo sapiens 358His Pro Ser Pro Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His1 5 10 15 Leu Thr Gly Lys Ser Asn 20 35920PRTHomo sapiens 359Leu Ile Ser Ser Tyr Gly Thr His Phe Ile Arg Ser Met Glu Leu

Gly1 5 10 15 Gly Arg Ile Ser 20 36018PRTHomo sapiens 360Gly Pro Val Gly Thr Glu Leu Phe Arg Val Pro Pro Val Ser Thr Ala1 5 10 15 Ala Thr36120PRTHomo sapiens 361Leu Thr Ser Ser Val Thr Ile Leu Pro Leu Pro Leu Gln Asn Ala Thr1 5 10 15 Val Glu Ala Gly 20 36222PRTHomo sapiens 362Leu Glu Arg Lys Ala Lys Trp Thr Thr Ala Val Arg Pro Leu Arg Leu1 5 10 15 Pro Ser Ser Lys Ala Gln 20 36323PRTHomo sapiens 363Gly Lys Lys Thr Pro Ala Lys Val Val Pro Met Lys Ala Lys Ser Val1 5 10 15 Ala Glu Glu Glu Asp Asp Glu 20 36421PRTHomo sapiens 364Lys Asp Gly Val Thr Pro Ile Lys Asp Leu Thr Ala His Phe Arg Gly1 5 10 15 Asp Arg Cys Lys Thr 20 36543PRTHomo sapiens 365Val Asn Trp Gly Arg Ile Val Ala Phe Phe Glu Phe Gly Gly Val Met1 5 10 15 Cys Val Glu Ser Val Asn Arg Glu Met Ser Pro Leu Val Asp Asn Ile 20 25 30 Ala Leu Trp Met Thr Glu Tyr Leu Asn Arg His 35 40 36643PRTHomo sapiens 366Pro Asn Trp Gly Arg Leu Val Ala Phe Phe Val Phe Gly Ala Ala Leu1 5 10 15 Cys Ala Glu Ser Val Asn Lys Glu Met Glu Pro Leu Val Gly Gln Val 20 25 30 Gln Glu Trp Met Val Ala Tyr Leu Glu Thr Arg 35 40 36743PRTHomo sapiens 367Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu1 5 10 15 Val Leu Lys Ala Leu Cys Thr Lys Val Pro Glu Leu Ile Arg Thr Ile 20 25 30 Met Gly Trp Thr Leu Asp Phe Leu Arg Glu Arg 35 40 36888PRTHomo sapiens 368Leu Arg Asp Ala His Gly Pro Pro Pro Ala Leu Val Phe Ser Cys Gln1 5 10 15 Met Gly Val Gly Arg Thr Asn Leu Gly Met Val Leu Gly Thr Leu Ile 20 25 30 Leu Leu His Arg Ser Gly Thr Thr Ser Gln Pro Glu Ala Ala Pro Thr 35 40 45 Gln Ala Lys Pro Leu Pro Met Glu Gln Phe Gln Val Ile Gln Ser Phe 50 55 60 Leu Arg Met Val Pro Gln Gly Arg Arg Met Val Glu Glu Val Asp Arg65 70 75 80 Ala Ile Thr Ala Cys Ala Glu Leu 85 36943PRTHomo sapiens 369Thr Cys Leu Thr Ile Val Gln Lys Leu Lys Lys Met Val Asp Lys Pro1 5 10 15 Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg Val Cys Arg Thr Gly 20 25 30 Arg Ser Arg Trp Arg Asp Val Cys Arg Asn Phe 35 40 37049PRTHomo sapiens 370Glu Arg Leu Ser Arg Pro Asn Ile Phe Ile Leu Asn Asn Arg Trp Asp1 5 10 15 Ala Ser Ala Ser Glu Pro Glu Tyr Met Glu Glu Val Arg Arg Gln His 20 25 30 Met Glu Arg Cys Thr Ser Phe Leu Val Asp Glu Leu Gly Val Val Asp 35 40 45 Arg 37146PRTHomo sapiens 371Cys Gln Phe Leu Ser Thr Phe Lys Glu Ser Cys Leu Gln Phe Tyr His1 5 10 15 Ala Glu Leu Lys Glu Leu Ser Phe Ile Arg Ala Ala Glu Glu Ser Arg 20 25 30 Lys His Ile Asn Thr Trp Val Ser Lys Lys Thr Glu Gly Lys 35 40 45 37251PRTHomo sapiens 372Leu Arg Asn Lys Leu Gln Ser Gln Leu Leu Ser Ile Glu Lys Glu Val1 5 10 15 Glu Glu Tyr Lys Asn Phe Arg Pro Asp Asp Pro Ala Arg Lys Thr Lys 20 25 30 Ala Leu Leu Gln Met Val Gln Gln Phe Ala Val Asp Phe Glu Lys Arg 35 40 45 Ile Glu Gly 50 37389PRTHomo sapiens 373Gly Pro Pro Val Ser Ser Ser Ser Asn Ser Ser Ser Ser Thr Ser Gln1 5 10 15 Gly Ser Tyr Pro Cys Ser His Pro Ser Pro Ser Gln Gly Pro Gln Gly 20 25 30 Ala Pro Tyr Pro Phe Pro Pro Val Pro Thr Val Thr Thr Ser Ser Ala 35 40 45 Thr Leu Ser Thr Val Ile Ala Thr Val Ala Ser Ser Pro Ala Gly Tyr 50 55 60 Lys Thr Ala Ser Pro Pro Gly Pro Pro Pro Tyr Gly Lys Arg Ala Pro65 70 75 80 Ser Pro Gly Ala Tyr Lys Thr Ala Ile 85 37444PRTHomo sapiens 374Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu1 5 10 15 Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr 20 25 30 Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe Arg 35 40 37546PRTHomo sapiens 375Ala Leu Thr Ala Leu Arg Thr Cys Glu Leu Ala Leu Glu Gly Leu Thr1 5 10 15 Ala Ser Glu Val Glu Asp Cys Leu Ala Val Glu Ala Glu Val Ser Ile 20 25 30 Ser Asp Arg Ala Ser Ala Ser Pro Ser Phe Lys Ala Cys Glu 35 40 45 37646PRTHomo sapiens 376Trp Gln Ser Ser Ala Pro His Gln Pro Gly Pro Ser Leu Trp Ala Glu1 5 10 15 Ala Lys Thr Ser Glu Ala Pro Ser Thr Gln Asp Pro Ser Thr Gln Ala 20 25 30 Ser Thr Ala Ser Ser Pro Ala Pro Glu Glu Asn Ala Pro Ser 35 40 45 37742PRTHomo sapiens 377Thr Arg Cys Gln Val Ala Gly Trp Gly Ser Gln Arg Ser Gly Gly Arg1 5 10 15 Leu Ser Arg Phe Pro Arg Phe Val Asn Val Thr Val Thr Pro Glu Asp 20 25 30 Gln Cys Arg Pro Asn Asn Val Cys Thr Gly 35 40 37858PRTHomo sapiens 378Arg Pro Leu Arg Leu Pro Ser Ser Lys Ala Gln Val Lys Pro Gly Gln1 5 10 15 Leu Cys Ser Val Ala Gly Trp Gly Tyr Val Ser Met Ser Thr Leu Ala 20 25 30 Thr Thr Leu Gln Glu Val Leu Leu Thr Val Gln Lys Asp Cys Gln Cys 35 40 45 Glu Arg Leu Phe His Gly Asn Tyr Ser Arg 50 55 37947PRTHomo sapiens 379Glu Glu Asp Glu Asp Asp Glu Asp Glu Asp Asp Glu Glu Glu Asp Asp1 5 10 15 Glu Asp Asp Asp Glu Glu Glu Glu Glu Glu Glu Pro Val Lys Ala Ala 20 25 30 Pro Gly Lys Arg Lys Lys Glu Met Thr Lys Gln Lys Glu Ala Pro 35 40 45 38045PRTHomo sapiens 380Leu Leu Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Arg Gly Thr1 5 10 15 Glu Leu Asp Asp Gly Ile Gln Ala Asp Ser Gly Pro Ile Asn Asp Thr 20 25 30 Asp Ala Asn Pro Arg Tyr Lys Ile Pro Val Glu Ala Asp 35 40 45 38141PRTHomo sapiens 381Gly Pro Ala Leu Ser Pro Val Pro Pro Val Val His Leu Thr Leu Arg1 5 10 15 Gln Ala Gly Asp Asp Phe Ser Arg Arg Tyr Arg Arg Asp Phe Ala Glu 20 25 30 Met Ser Ser Gln Leu His Leu Thr Pro 35 40 38231PRTHomo sapiens 382Leu His Gln Ala Met Arg Ala Ala Gly Asp Glu Phe Glu Thr Arg Phe1 5 10 15 Arg Arg Thr Phe Ser Asp Leu Ala Ala Gln Leu His Val Thr Pro 20 25 30 38331PRTHomo sapiens 383Thr Lys Lys Leu Ser Glu Cys Leu Lys Arg Ile Gly Asp Glu Leu Asp1 5 10 15 Ser Asn Met Glu Leu Gln Arg Met Ile Ala Ala Val Asp Thr Asp 20 25 30 38458PRTHomo sapiens 384Val Ser Tyr Thr Pro Arg Asp Lys Gln Asn Leu His Glu Asn Leu Gln1 5 10 15 Gly Leu Gly Pro Gly Val Arg Val Glu Ser Leu Glu Leu Ala Ile Arg 20 25 30 Lys Glu Ile His Asp Phe Ala Gln Leu Ser Glu Asn Thr Tyr His Val 35 40 45 Tyr His Asn Thr Glu Asp Leu Trp Gly Glu 50 55 38529PRTHomo sapiens 385Leu Val Phe Ser Arg Leu Ser Pro Glu Tyr Tyr Asp Pro Ala Arg Ala1 5 10 15 His Leu Arg Asp Gly Glu Lys Ser Cys Pro Cys Gly Gln 20 25 38644PRTHomo sapiens 386Phe Lys Gly Lys Trp Asn Glu Pro Phe Asp Glu Thr Tyr Thr Arg Glu1 5 10 15 Met Pro Phe Lys Ile Asn Gln Glu Glu Gln Arg Pro Val Gln Met Met 20 25 30 Tyr Gln Glu Ala Thr Phe Lys Leu Ala His Val Gly 35 40 38748PRTHomo sapiens 387Tyr Arg Phe Ser Met Asp Leu Val Glu Cys Arg Phe Tyr Ser Phe His1 5 10 15 Leu Val His Thr Pro Pro Val His Pro Glu Phe Lys Arg Ala Leu Lys 20 25 30 Thr Leu Pro Pro His Phe Asn Thr Ser Thr Lys Pro Asp Tyr His Arg 35 40 45 38839PRTHomo sapiens 388Ile Ser Ser Met Ser Glu Asn Gly Tyr Asp Pro Gln Gln Asn Leu Asn1 5 10 15 Asp Leu Met Leu Leu Gln Leu Asp Arg Glu Ala Asn Leu Thr Ser Ser 20 25 30 Val Thr Ile Leu Pro Leu Pro 35 38940PRTHomo sapiens 389Val Lys Arg Pro Ile Pro His Pro Ala Tyr Asn Pro Lys Asn Phe Ser1 5 10 15 Asn Asp Ile Met Leu Leu Gln Leu Glu Arg Lys Ala Lys Trp Thr Thr 20 25 30 Ala Val Arg Pro Leu Arg Leu Pro 35 40 39048PRTHomo sapiens 390Gly Thr Ser Pro Glu Leu Pro Thr Gly Val Thr Gly Ser Ser Gly Thr1 5 10 15 Arg Leu Pro Pro Thr Pro Lys Ala Gln Asp Gly Gly Pro Val Gly Thr 20 25 30 Glu Leu Phe Arg Val Pro Pro Val Ser Thr Ala Ala Thr Trp Gln Ser 35 40 45 39140PRTHomo sapiens 391Asp Cys Ser Cys Ala Lys Met Gln Asp Leu Leu Lys Lys Ala Ser Glu1 5 10 15 Glu Asp His Thr Asn Ala Ala Cys Phe Ala Cys Ile Leu Leu Ser His 20 25 30 Gly Glu Glu Asn Val Ile Tyr Gly 35 40 39237PRTHomo sapiens 392Leu Gly Met Phe Gln Leu Phe His Leu Gln Lys Glu Leu Ala Glu Leu1 5 10 15 Arg Glu Ser Thr Ser Gln Met His Thr Ala Ser Ser Leu Glu Lys Gln 20 25 30 Ile Gly His Pro Ser 35 39318PRTHomo sapiens 393Phe Thr Ala Arg Gly Arg Phe Ala Thr Val Val Glu Glu Leu Phe Arg1 5 10 15 Asp Gly39418PRTHomo sapiens 394Gly Ser Ala Gln Gln Arg Phe Thr Gln Val Ser Asp Glu Leu Phe Gln1 5 10 15 Gly Gly39518PRTHomo sapiens 395Ser Pro Arg Glu Val Phe Phe Arg Val Ala Ala Asp Met Phe Ser Asp1 5 10 15 Gly Asn39655PRTHomo sapiens 396Phe Leu Gln Pro Thr Tyr Arg Tyr His Arg Leu Pro Leu Pro Glu Gln1 5 10 15 Gly Ser Pro Leu Glu Ala Gln Leu Asp Ala Phe Val Ser Val Leu Arg 20 25 30 Glu Thr Pro Ser Leu Leu Gln Leu Arg Asp Ala His Gly Pro Pro Pro 35 40 45 Ala Leu Val Phe Ser Cys Gln 50 55 39719PRTHomo sapiens 397Glu Gly Pro Gln Gly Asp Leu Leu Thr Lys Thr Gln Glu Leu Gly Arg1 5 10 15 Asp Tyr Arg39826PRTHomo sapiens 398Glu Val Arg Ala Gln Leu Leu Glu Leu Pro Tyr Ala Arg Lys Glu Leu1 5 10 15 Ser Leu Leu Val Leu Leu Pro Asp Asp Gly 20 25 39920PRTHomo sapiens 399Leu Ile Ser Ser Tyr Gly Thr His Phe Ile Arg Ser Met Glu Leu Gly1 5 10 15 Gly Arg Ile Ser 20 40020PRTHomo sapiens 400Leu Gln Asn Ala Thr Val Glu Ala Gly Thr Arg Cys Gln Val Ala Gly1 5 10 15 Trp Gly Ser Gln 20 40120PRTHomo sapiens 401Ser Ser Lys Ala Gln Val Lys Pro Gly Gln Leu Cys Ser Val Ala Gly1 5 10 15 Trp Gly Tyr Val 20 40220PRTHomo sapiens 402Ser Ala Pro His Gln Pro Gly Pro Ser Leu Trp Ala Glu Ala Lys Thr1 5 10 15 Ser Glu Ala Pro 20 40321PRTHomo sapiens 403Lys Asp Gly Val Thr Pro Ile Lys Asp Leu Thr Ala His Phe Arg Gly1 5 10 15 Asp Arg Cys Lys Thr 20 40419PRTHomo sapiens 404Pro Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His Leu Thr Gly1 5 10 15 Lys Ser Asn40542PRTHomo sapiens 405Val Asn Trp Gly Arg Ile Val Ala Phe Phe Glu Phe Gly Gly Val Met1 5 10 15 Cys Val Glu Ser Val Asn Arg Glu Met Ser Pro Leu Val Asp Asn Ile 20 25 30 Ala Leu Trp Met Thr Glu Tyr Leu Asn Arg 35 40 40643PRTHomo sapiens 406Pro Asn Trp Gly Arg Leu Val Ala Phe Phe Val Phe Gly Ala Ala Leu1 5 10 15 Cys Ala Glu Ser Val Asn Lys Glu Met Glu Pro Leu Val Gly Gln Val 20 25 30 Gln Glu Trp Met Val Ala Tyr Leu Glu Thr Arg 35 40 40743PRTHomo sapiens 407Phe Asn Trp Gly Arg Val Val Ala Leu Phe Tyr Phe Ala Ser Lys Leu1 5 10 15 Val Leu Lys Ala Leu Cys Thr Lys Val Pro Glu Leu Ile Arg Thr Ile 20 25 30 Met Gly Trp Thr Leu Asp Phe Leu Arg Glu Arg 35 40 40872PRTHomo sapiens 408Ala Leu Val Phe Ser Cys Gln Met Gly Val Gly Arg Thr Asn Leu Gly1 5 10 15 Met Val Leu Gly Thr Leu Ile Leu Leu His Arg Ser Gly Thr Thr Ser 20 25 30 Gln Pro Glu Ala Ala Pro Thr Gln Ala Lys Pro Leu Pro Met Glu Gln 35 40 45 Phe Gln Val Ile Gln Ser Phe Leu Arg Met Val Pro Gln Gly Arg Arg 50 55 60 Met Val Glu Glu Val Asp Arg Ala65 70 40943PRTHomo sapiens 409Thr Cys Leu Thr Ile Val Gln Lys Leu Lys Lys Met Val Asp Lys Pro1 5 10 15 Thr Gln Arg Ser Val Ser Asn Ala Ala Thr Arg Val Cys Arg Thr Gly 20 25 30 Arg Ser Arg Trp Arg Asp Val Cys Arg Asn Phe 35 40 41053PRTHomo sapiens 410Val Glu Leu Ser Thr Val Glu Lys Ser Leu Thr Phe Glu Lys Leu Thr1 5 10 15 Ala Trp Thr Lys Pro Asp Cys Met Lys Ser Thr Glu Val Glu Leu Val 20 25 30 Leu Leu Pro Lys Phe Lys Leu Gln Glu Asp Tyr Asp Met Glu Ser Val 35 40 45 Leu Arg His Leu Gly 50 41155PRTHomo sapiens 411Ala Leu Thr Ala Leu Arg Thr Cys Glu Leu Ala Leu Glu Gly Leu Thr1 5 10 15 Ala Ser Glu Val Glu Asp Cys Leu Ala Val Glu Ala Glu Val Ser Ile 20 25 30 Ser Asp Arg Ala Ser Ala Ser Pro Ser Phe Lys Ala Cys Glu Glu Lys 35 40 45 Lys Lys Asn His Lys Val Gly 50 55 41243PRTHomo sapiens 412Arg Ser Gly Gly Arg Leu Ser Arg Phe Pro Arg Phe Val Asn Val Thr1 5 10 15 Val Thr Pro Glu Asp Gln Cys Arg Pro Asn Asn Val Cys Thr Gly Val 20 25 30 Leu Thr Arg Arg Gly Gly Ile Cys Asn Gly Asp 35 40 41349PRTHomo sapiens 413Ser Met Ser Thr Leu Ala Thr Thr Leu Gln Glu Val Leu Leu Thr Val1 5 10 15 Gln Lys Asp Cys Gln Cys Glu Arg Leu Phe His Gly Asn Tyr Ser Arg 20 25 30 Ala Thr Glu Ile Cys Val Gly Asp Pro Lys Lys Thr Gln Thr Gly Phe 35 40 45 Lys 41452PRTHomo sapiens 414Ser Thr Gln Asp Pro Ser Thr Gln Ala Ser Thr Ala Ser Ser Pro

Ala1 5 10 15 Pro Glu Glu Asn Ala Pro Ser Glu Gly Gln Arg Val Trp Gly Gln Gly 20 25 30 Gln Ser Pro Arg Glu Asn Ser Leu Glu Arg Glu Glu Met Gly Pro Val 35 40 45 Pro Ala His Thr 50 41551PRTHomo sapiens 415Leu Leu Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Arg Gly Thr1 5 10 15 Glu Leu Asp Asp Gly Ile Gln Ala Asp Ser Gly Pro Ile Asn Asp Thr 20 25 30 Asp Ala Asn Pro Arg Tyr Lys Ile Pro Val Glu Ala Asp Phe Leu Phe 35 40 45 Ala Tyr Ser 50 41652PRTHomo sapiens 416Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu1 5 10 15 Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr 20 25 30 Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys 35 40 45 Asn Asn Leu Pro 50 41747PRTHomo sapiens 417Gly Gly Trp Asp Ala Phe Val Glu Leu Tyr Gly Pro Ser Met Arg Pro1 5 10 15 Leu Phe Asp Phe Ser Trp Leu Ser Leu Lys Thr Leu Leu Ser Leu Ala 20 25 30 Leu Val Gly Ala Cys Ile Thr Leu Gly Ala Tyr Leu Gly His Lys 35 40 45 41852PRTHomo sapiens 418Gly Gly Trp Ala Glu Phe Thr Ala Leu Tyr Gly Asp Gly Ala Leu Glu1 5 10 15 Glu Ala Arg Arg Leu Arg Glu Gly Asn Trp Ala Ser Val Arg Thr Val 20 25 30 Leu Thr Gly Ala Val Ala Leu Gly Ala Leu Val Thr Val Gly Ala Phe 35 40 45 Phe Ala Ser Lys 50 41963PRTHomo sapiens 419Gly Gly Trp Val Arg Leu Leu Lys Pro Pro His Pro His His Arg Ala1 5 10 15 Leu Thr Thr Ala Pro Ala Pro Pro Ser Leu Pro Pro Ala Thr Pro Leu 20 25 30 Gly Pro Trp Ala Phe Trp Ser Arg Ser Gln Trp Cys Pro Leu Pro Ile 35 40 45 Phe Arg Ser Ser Asp Val Val Tyr Asn Ala Phe Ser Leu Arg Val 50 55 60 42070PRTHomo sapiens 420Ala Gly Pro Val Ala Pro Arg Asp Leu Ile Ala Arg Gly Ser Leu Arg1 5 10 15 Glu Asp Asp Leu Val Ser Pro Asp Ala Leu Ser Thr Val Arg Glu Met 20 25 30 Asp Val Ala Asn Phe Arg Arg Val Pro Arg Met Pro Ile Tyr Gly Thr 35 40 45 Ala Gln Pro Ser Ala Lys Ala Leu Gly Ser Ile Leu Ala Tyr Leu Thr 50 55 60 Asp Ala Lys Arg Arg Ile65 70 42135PRTHomo sapiens 421Met Arg Arg Tyr Gln Ser Arg Val Ile Gln Gly Leu Val Ala Gly Glu1 5 10 15 Thr Ala Gln Gln Ile Cys Glu Asp Leu Arg Leu Cys Ile Pro Ser Thr 20 25 30 Gly Pro Leu 35 42254PRTHomo sapiens 422Ala Asp Leu Ser Ala Met Ser Ala Glu Arg Asp Leu Cys Leu Ser Lys1 5 10 15 Phe Val His Lys Ser Phe Val Glu Val Asn Glu Glu Gly Thr Glu Ala 20 25 30 Ala Ala Ala Ser Ser Cys Phe Val Val Ala Glu Cys Cys Met Glu Ser 35 40 45 Gly Pro Arg Phe Cys Ala 50 42354PRTHomo sapiens 423Glu Arg His Ser Asn Val Asp Gly Gly His His Ser Thr Met His Asp1 5 10 15 Leu Leu Phe Gly Ser Gln Ala Gly Pro Glu Gln Phe Ser Ala Trp Val 20 25 30 Ala Ser Leu Gln Asp Ser Pro Gly Leu Val Asp Tyr Thr Leu Glu Pro 35 40 45 Leu His Met Leu Val Glu 50 42451PRTHomo sapiens 424Gly Gly Thr Pro Leu Val Cys Glu Gly Leu Ala His Gly Val Ala Ser1 5 10 15 Phe Ser Leu Gly Pro Cys Gly Arg Gly Pro Asp Phe Phe Thr Arg Val 20 25 30 Ala Leu Phe Arg Asp Trp Ile Asp Gly Val Leu Asn Asn Pro Gly Pro 35 40 45 Gly Pro Ala 50 42547PRTHomo sapiens 425Gly Asp Ser Gly Gly Pro Leu Val Cys Lys Asp Val Ala Gln Gly Ile1 5 10 15 Leu Ser Tyr Gly Asn Lys Lys Gly Thr Pro Pro Gly Val Tyr Ile Lys 20 25 30 Val Ser His Phe Leu Pro Trp Ile Lys Arg Thr Met Lys Arg Leu 35 40 45 42657PRTHomo sapiens 426Met Ala His Val Ser Val Val Pro Val Ser Ser Glu Gly Thr Pro Ser1 5 10 15 Arg Glu Pro Val Ala Ser Gly Ser Trp Thr Pro Lys Ala Glu Glu Pro 20 25 30 Ile His Ala Thr Met Asp Pro Gln Arg Leu Gly Val Leu Ile Thr Pro 35 40 45 Val Pro Asp Ala Gln Ala Ala Thr Arg 50 55 42754PRTHomo sapiens 427Thr Val Pro Gly Tyr Tyr Ser Trp Arg Ser Pro Gly Arg Gly Ser Trp1 5 10 15 Phe Val Gln Ala Leu Cys Ser Ile Leu Glu Glu His Gly Lys Asp Leu 20 25 30 Glu Ile Met Gln Ile Leu Thr Arg Val Asn Asp Arg Val Ala Arg His 35 40 45 Phe Glu Ser Gln Ser Asp 50 42853PRTHomo sapiens 428Leu Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Gln Asp Leu1 5 10 15 Val Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr Gly Met Trp 20 25 30 Ala Arg Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 35 40 45 His Leu Tyr Val Asn 50

Claims

1. An isolated peptide comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 17, 18, 19, 21-25, 30, 31-36, 39-47, 49-52, 54-57, 59-63, 66-75, 84-93, 102-106, 108-121, 132-175, 179-187, 191-199, 205-209, 211-223, 227-235, 238-243, 245-247, 249-251, 253-256 and 260-263, wherein the amino acid residue represented by (x) is a serine, a threonine, a tryptophan, a H-bond donor residue or a H-bond acceptor residue, wherein the amino acid residue represented by (b) is a lysine, an arginine, an asparagine, a glutamine or a basic residue, wherein the amino acid residue represented by (j) is a cysteine or a thiol residue, wherein in the amino acid residue represented by (o) is an anthrylalanine or other non-natural amino acid and wherein said peptide induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity.

2-170. (canceled)

171. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 17, 18, 19, 21-25, 269 and 272.

172. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 30, 31-36 and 273.

173. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 39-47.

174. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 49-52.

175. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 54-57.

176. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 59-63 and 274.

177. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 66-75.

178. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 84-93 and 275.

179. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 102-106, 108-121, 267, 276 and 277.

180. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 132-175.

181. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 179-187, 266 and 278.

182. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NO: 191-199.

183. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 205-209 and 211-223.

184. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 227-235.

185. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 238-243, 245-247, 265 and 279.

186. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 249-251.

187. The isolated protein of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 253-256.

188. The isolated peptide of claim 1 comprising one or more amino acid sequence selected from the group consisting of SEQ ID NOS: 260-263 and 268.

189. An isolated peptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NOS: 14-16, 20, 26-29, 37, 38, 48, 53, 58, 64, 65, 72, 76-83, 94-101, 107, 114, 122-131, 170, 176-178, 188-190, 200-204, 210, 224-226, 236, 237, 244, 248, 252, 257-259 and 288-289, wherein said peptide induces antimicrobial, anti-cancer, anti-inflammatory, anti-proliferative or programmed cell death activity.

190. A method of inducing programmed cell death in a cell, comprising contacting said cell with the isolated peptide of claim 1.

191. A method of inducing programmed cell death in a cell, comprising contacting said cell with the isolated peptide of claim 189.

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