ATP binding site of mitochondrial creatine kinase

FEBS Letters

James, Peter²  Wallimann, Theo¹  Carafoli, Ernesto²  Lutsenko, Svetlana³  Wyss, Markus¹ 

[1] Institute for Cell Biology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland;Laboratory of Biochemistry, Swiss Federal Institute of Technology, ETH-Zentrum, CH-8092 Zürich, Switzerland;Shemyakin Institute of Bioorganic Chemistry, USSR Academy of Sciences, UI. Miklukho-Maklaya 16/10, 117871 GSP Moscow, V-437 USSR

关键词: ATP binding site;    Creatine kinase;    CIRATP analogue;    CK;    creatine kinase;    Mib-CK;    mitochondrial isoen-zyme of creatine kinase from chicken heart;    CIRATP;    γ-[4-(N-2-chloroethyl-A'-methylamino)]benzylamide adenosine triphosphate;    BSA;    bovine serum albumin;   

DOI  :  10.1016/0014-5793(90)81069-Z

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The ATP binding site of mitochondrial creatine kinase from chicken heart has been studied by modifying the purified enzyme with a ¹⁴C-labelled ATP analogue, CIRATP, in which the reactive label was covalently bound to the γ-phosphote group of ATP. The modified enzyme was digested by pepsin, and a single radioactive nonapeptide was isolated by HPLC. Amino acid analysis and direct sequence determination revealed that the isolated peptide corresponds to amino acids 335–343 within the C-terminal region of Mi-CK, this peptide being highly preserved throughout evolution. Asp-335 is very likely the site of modification by CIRATP. The specificity of the ATP analogue for the active site of creatine kinase was demonstrated by the inhibition of the enzymatic activity of Mi-CK by C1RATP and by the prevention of this inhibition bij ADP.

【 授权许可】

Unknown