A sensitive, simple assay of mitochondrial ATP synthesis of cultured mammalian cells suitable for high-throughput analysis

被引：28

作者：

Fujikawa, Makoto ^{[2
]}

Yoshida, Masasuke ^{[1
,2
]}

机构：

[1] Kyoto Sangyo Univ, Dept Mol Biosci, Fac Life Sci, Kyoto 6038555, Japan

[2] Japan Sci & Technol Agcy JST, ICORP ATP Synth Regulat Project, Tokyo 1350064, Japan

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2010年 / 401卷 / 04期

关键词：

FoF1 ATP synthase; ATP synthesis; Mitochondria; High-throughput analysis; Streptolysin; STREPTOLYSIN-O; IDENTIFICATION; SYNTHASE;

D O I：

10.1016/j.bbrc.2010.09.089

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

A new assay has been developed to measure mitochondrial ATP synthesis of cultured mammalian cells Cells in a microplate are exposed to streptolysin O to make plasma membranes permeable without damaging mitochondrial function and ATP synthesis is monitored by luciferase Addition of inhibitors of FoF1-ATP synthase (FoF1) respiratory chain TCA cycle and ATP/ADP translocator as well as knockdown of beta-subunit of FoF(1) resulted in loss of ATP synthesis Compared with the conventional procedures that need mitochondria fractionation and detergent this assay is simple sensitive and suitable for high-throughput analysis of genes and drugs that could affect mitochondrial functional integrity as represented by ATP synthesis activity (C) 2010 Elsevier Inc All rights reserved

引用

页码：538 / 543

页数：6

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