title: Structure-based design of an agonistic peptide targeting Fas.
abstract: A small agonistic peptide FRAP-4 (WEWT, Fas reactive peptide-4) that binds to the human Fas molecule was discovered using our computer screening strategy named the Amino acid Complement Wave (ACW) method, which is based on the complementarities of interacting amino acids between comprehensive testing peptides and a target protein surface pocket. In silico docking studies demonstrated the specific interaction of FRAP-4 with the main Fas ligand (FasL) binding domain in the Fas molecule. An octamer of this peptide produced by carboxyl terminal linkages of polylysine branches (MAP), (FRAP-4)8-MAP, effectively induced apoptosis in human ovarian cancer cell line NOS4 cells that was associated with the activation of caspases-8, -9 and -3, and the cleavage of PARP. Alanine substitution of the N-terminal W in FRAP-4 resulted in complete loss of FasL-mimetic action of (FRAP-4)8-MAP, suggesting that the aromatic functionality at the N-terminal position W appears to play an essentially important role in Fas binding ability. These observations indicate that the FasL-mimetic peptide should serve as an excellent starting point for the design of effective compounds with FasL-mimetic activity. Furthermore, the ACW method for the structure-based design of optimized small peptides against receptor molecules such as Fas could open new avenues for the development of peptide mimetic and nonpeptidic organic forms to generate novel effective pharmaceuticals.
authors.affiliation: Research Institute for Biological Sciences, Tokyo University of Science, 2669 Yamazaki Noda, Chiba, 278-0022, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Yoshimori
authors.forename: R
authors.initials: R
authors.lastname: Takasawa
authors.forename: A
authors.initials: A
authors.lastname: Hayakawa
authors.forename: M
authors.initials: M
authors.lastname: Mizuno
authors.forename: J
authors.initials: J
authors.lastname: Yoshida
authors.forename: S
authors.initials: S
authors.lastname: Tanuma
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008958:Models, Molecular; D015233:Models, Statistical; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011419:Propidium / Q000494:pharmacology; D011485:Protein Binding; D011487:Protein Conformation; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D012984:Software; D019014:fas Receptor / Q000378:metabolism*
chemical_list: D000596:Amino Acids; C505377:FRAP-4 peptide; D009363:Neoplasm Proteins; D010455:Peptides; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D018124:Receptors, Tumor Necrosis Factor; C105190:TNFRSF10A protein, human; D019014:fas Receptor; C030193:immunosuppressive acidic protein; D011419:Propidium; D011065:Poly(ADP-ribose) Polymerases; C505407:CASP3 protein, human; C505530:CASP8 protein, human; C506433:CASP9 protein, human; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases; D004229:Dithiothreitol