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. 2009 Oct:Chapter 19:Unit19.3.
doi: 10.1002/0471142905.hg1903s63.

Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using polarography and spectrophotometric enzyme assays

Antoni Barrientos  1 , Flavia Fontanesi, Francisca Díaz
Affiliations

Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using polarography and spectrophotometric enzyme assays

Antoni Barrientos et al. Curr Protoc Hum Genet. 2009 Oct.

Abstract

The oxidative phosphorylation (OXPHOS) system consists of five multimeric complexes embedded in the mitochondrial inner membrane. They work in concert to drive the aerobic synthesis of ATP. Mitochondrial and nuclear DNA mutations affecting the accumulation and function of these enzymes are the most common cause of mitochondrial diseases and have also been associated with neurodegeneration and aging. For this reason, several approaches for the assessment of the OXPHOS system enzymes have been developed. Based on the methods described elsewhere, the assays describe methods that form a biochemical characterization of the OXPHOS system in cells and mitochondria isolated from cultured cells or tissues.

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Figures

Figure 1
Figure 1
Polarographic assessment of mitochondrial respiratory chain performance. (A) Polarographic chamber (Hansatech). (B) and (C) Typical oxygen consumption recording chart using mitochondria isolated from either wild type cells or cells with partial respiratory deficiency, respectively. Pyruvate plus malate were used as site I substrates.
See this image and copyright information in PMC

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