In situ assay of the intramitochondrial enzymes: use of alamethicin for permeabilization of mitochondria

被引：78

作者：

Gostimskaya, IS

Grivennikova, VG

Zharova, TV

Bakeeva, LE

Vinogradov, AD ^{[1
]}

机构：

[1] Moscow MV Lomonosov State Univ, Sch Biol, Dept Biochem, Moscow 119992, Russia

[2] Moscow MV Lomonosov State Univ, AN Belozersky Inst Phys Chem Biol, Moscow 119992, Russia

来源：

ANALYTICAL BIOCHEMISTRY | 2003年 / 313卷 / 01期

关键词：

intramitochondrial enzymes; membrane permeabilization; alamethicin; complex I; F center dot F-1 -ATPase;

D O I：

10.1016/S0003-2697(02)00534-1

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The channel-forming antibiotic alamethicin was used to permeabilize mitochondrial membranes for the low molecular mass hydrophilic substrates NADH and ATP. Alamethicin-treated mitochondria show high rotenone-sensitive NADH oxidase, NADH-quinone reductase, and oligomycin-sensitive and carboxyatractylate-insensitive ATPase activities. Alamethicin does not affect Complex I and ATPase activities in inside-out submitochondrial particles. Permeabilized mitochondria quantitatively retain their aconitase and iso-citrate dehydrogenase activities. Electron microscopy of alamethicin-treated mitochondria reveals no disruption of their outer and inner membranes. From the results obtained it is recommended, that alamethicin be used for the in situ catalytic assay of intramitochondrially located enzymes. (C) 2003 Elsevier Science (USA). All rights reserved.

引用

页码：46 / 52

页数：7

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