Mapping of cyclosporin A binding sites in cyclophilin A by using synthetic peptides.
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 alpha-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.
Yamamoto
K
K
Laboratory of Pharmaceutical Sciences, Osaka University School of Medicine, 2-15 Yamadaoka, Suita-shi, Osaka-fu 565, Japan.
Kurokawa
N
N
Kadobayashi
M
M
Tauchi
N
N
Iguchi
K
K
Yanaihara
N
N
Yanaihara
C
C
D000595:Amino Acid Sequence; D001665:Binding Sites; D021984:Cyclophilin A / Q000737:chemistry* / Q000378:metabolism*; D016572:Cyclosporine / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism
D010446:Peptide Fragments; D016572:Cyclosporine; D021984:Cyclophilin A