Assay of mitochondrial respiratory chain complex I in human lymphocytes and cultured skin fibroblasts

被引：48

作者：

Chretien, D ^{[1
]}

Bénit, P ^{[1
]}

Chol, M ^{[1
]}

Lebon, S ^{[1
]}

Rötig, A ^{[1
]}

Munnich, A ^{[1
]}

Rustin, P ^{[1
]}

机构：

[1] Hop Necker Enfants Malad, INSERM, Unite Rech Handicaps Genet Enfant, U393, F-75743 Paris 15, France

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2003年 / 301卷 / 01期

关键词：

human; mitochondria; respiratory chain; complex I assay; lymphocytes; fibroblasts;

D O I：

10.1016/S0006-291X(02)03016-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

Respiratory chain complex I (NADH:ubiquinone oxidoreductase) deficiency is one of the most frequent causes of mitochondrial disease in humans. The activity of this complex can be confidently measured in most tissue samples, but not in cultured skin fibroblasts or circulating lymphocytes. Highly contaminating non-mitochondrial NADH-quinone oxidoreductase activity in fibroblasts and the limited access of substrates to complex I in lymphocytes hinder its measurement in permeabilized cells. Complex I assay in these cells requires the isolation of mitochondria, which in turn necessitates large quantities of cells and is not feasible when studying circulating lymphocytes. Here we report a simple method to measure complex I activity in a minute amount of either cell type. The procedure strongly reduces contaminating NADH:quinone oxidoreductase activity and permits measuring high rates of rotenone-sensitive complex I activity thanks to effective cell permeabilization. (C) 2002 Elsevier Science (USA). All rights reserved.

引用

页码：222 / 224

页数：3

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