Identification of a cross‐linked double‐peptide from the catalytic site of the Ca2+‐ATPase of sarcoplasmic reticulum formed by the Ca2+‐ and pH‐dependent reaction with ATP γP‐imidazolidate

FEBS Letters

Mann, Karlheinz¹  Bäumert, Hans G.²  Gutowski-Eckel, Zeynep² 

[1] Max-Planck-Institut für Biochemie, Martinsried, Germany;Institut für Biophysikalische Chemie und Biochemie der Johann-Wolfgang-Goethe Universität, Haus 75A, Klinikum, Theodor-Stern-Kai 7, D-6000 Frankfurt/Main 70, Germany

关键词: Ca2+-ATPase;    Sarcoplasmic reticulum;    ATP γP-imidazolidate;    Intramolecular cross-linking;    Double-peptide;    Catalytic site;    SR;    sarcoplasmic reticulum;    AP3PL;    adenosine 5'-triphosphopyridoxal;    TNP-8-N3;    -ATP;    2';    3'-O-(2;    4;    6-trinitrophenyl)-8-azido-ATP;   

DOI  :  10.1016/0014-5793(93)80142-H

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The Ca²⁺-ATPase from sarcoplasmic reticulum can be inhibited by the Ca²⁺- and pH-dependent reaction with ATP γP-imidazolidate. The chemically and monofunctionally activated inhibitor introduces an intramolecular cross-link between two neighbouring peptides of the active site. This can be followed by the reduced mobility of the ATPase upon SDS-PAGE analysis which becomes even more pronounced after limited trypsinolysis. After cleavage of the cross-linked ATPase molecule by cyanogen bromide and separation of the peptides a double-peptide can be detected which upon sequencing can be identified as part of the phosphorylation and the nucleotide binding site, respectively.

【 授权许可】

Unknown