Mapping of cyclosporin A binding sites in cyclophilin A by using synthetic peptides

• Published in: Regulatory peptides 1995-09, Vol.59 (1), p.23-30
• Main Authors: Yamamoto, Katsumi, Kurokawa, Nobuo, Kadobayashi, Muneo, Tauchi, Noriko, Iguchi, Kazuaki, Yanaihara, Noboru, Yanaihara, Chizuko
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Binding Sites; Biological and medical sciences; Cyclophilin A; Cyclophilin A - chemistry; Cyclophilin A - metabolism; Cyclosporin A; Cyclosporin A binding protein; Cyclosporin A binding site mapping; Cyclosporine - metabolism; Humans; Immunomodulators; Medical sciences; Molecular Sequence Data; Peptide Fragments - chemical synthesis; Peptide Fragments - chemistry; Peptide Fragments - metabolism; Pharmacology. Drug treatments; Simultaneous multiple peptide synthesis
• ISSN: 0167-0115; 1873-1686
• DOI: 10.1016/0167-0115(95)00067-L

In order to map cyclosporin A (CsA) binding sites of cyclophilin (CyP), we synthesized the complete set of overlapping 157 octapeptides corresponding to human CyP A using the multi-pin peptide synthesis system. The pin-coupled synthetic octapeptides were examined in terms of binding ability to CsA by a modification of the enzyme-linked immunosorbent assay. Significant binding of CsA was detected with 35 synthetic N α-acetylated octapeptides possessing the N-terminal amino acids corresponding to the residues in positions 24–26, 42–44, 69–73, 75, 76, 89–91, 102, 116, 124–131, 144–151 and 152 in human CyP A, respectively. Other eight octapeptides showed moderate CsA binding activity. The distinct binding of octapeptides covering the C-terminal region of the CyP A was particularly significant. These data are to be compared with the information provided by X-ray and NMR studies on the CsA binding sites and furnish thus a test of the reported method. The present study also gave added insight into the CsA interaction sites of CyP.