Detection of intermediary Clr with complete active site, using a synthetic proteinase inhibitor

FEBS Letters

Fujii, Setsuro¹  Ueno, Yoshihiko¹  Niinobe, Michio¹  Hitomi, Yuji¹ 

[1] Division of Regulation of Macromolecular Function, Institute for Protein Research, Osaka University, Suita, Osaka 565, Japan

关键词: Synthetic proteinase inhibitor;    6-Amidino-2-naphthyl-4-guanidinobenzoate;    FUT-175;    Clr activation;    Intermediary Clr;    Clr active site determination;    FUT-175;    6-amidino-2-naphthyl-4-gua-nidinobenzoate;    NPGB;    p-nitrophenyl-p'-guanidino-benzoate;    DFP;    diisopropyl fluorophosphate;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(84)81117-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The synthetic proteinase inhibitor, FUT-175 (6-amidino-2-naphthyl-4-guanidinobenzoate), strongly suppressed activation of Clr at 37°C, causing 50% inhibition at 0.03 mM. To clarify whether the inhibitor was incorporated into the active site of intermediary Clr formed during the incubation, determination of the active site was tried using this inhibitor. Consequently, release of amidinonaphthol equimolar with the amount of Clr used was observed in the early period of incubation, in which the activation to Clr was about 5%. These results indicate that intermediary Clr already has a complete active site.

【 授权许可】

Unknown