A sensitive, simple assay of mitochondrial ATP synthesis of cultured mammalian cells suitable for high-throughput analysis
A sensitive, simple assay of mitochondrial ATP synthesis of cultured mammalian cells suitable for high-throughput analysis
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DOI：
10.1016/j.bbrc.2010.09.089
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发表时间：
2010-10-29
影响因子：
3.1
通讯作者：
Yoshida, Masasuke
中科院分区：
文献类型：
--
作者：
Fujikawa, Makoto;Yoshida, Masasuke
一种新的测定培养的哺乳动物细胞的线粒体ATP合成的方法已经被开发出来，将微板中的细胞暴露于链球菌溶血素O以使质膜可渗透而不损害线粒体功能，并且通过荧光素酶监测ATP合成。添加FoF 1-ATP合成酶（FoF 1）呼吸链TCA循环和ATP/ADP转运体的抑制剂以及敲低FoF的β亚基（1）。与需要线粒体分离和去污剂的常规方法相比，该方法简单、灵敏，适用于以ATP合成活性为代表的影响线粒体功能完整性的基因和药物的高通量分析（C）2010 Elsevier Inc版权所有
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