磷酸化/去磷酸化测定

Phosphorylation/Dephosphorylation Assays.

作者信息

Suzuki Hiroshi

机构信息

Asahikawa Medical University, 2-1-1-1, Midorigaokahigashi, Asahikawa, Hokkaido, 078-8510, Japan.

出版信息

Methods Mol Biol. 2016;1377:211-26. doi: 10.1007/978-1-4939-3179-8_20.

DOI:10.1007/978-1-4939-3179-8_20

PMID:26695035

Abstract

The P-type ATPases form an autophosphorylated intermediate with ATP, and its isomeric transition and hydrolysis are obligatory events in the ATP-driven pump and thus for the energy coupling. The analyses of these reactions are therefore crucial for understanding the mechanism of the pump function and diseases caused by its defects. Here we describe the methods to analyze these processes in the transport cycle with a representative member of P-type ATPase family, SERCA1a, sarco(endo)plasmic reticulum Ca(2+)-ATPase.

摘要

P型ATP酶与ATP形成一种自磷酸化中间体，其异构化转变和水解是ATP驱动泵中必不可少的事件，因此对于能量偶联也是如此。因此，对这些反应的分析对于理解泵功能的机制及其缺陷所导致的疾病至关重要。在这里，我们描述了用P型ATP酶家族的一个代表性成员——肌浆（内质）网Ca²⁺-ATP酶（SERCA1a）来分析转运循环中这些过程的方法。