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. 2009 Oct:Chapter 19:Unit19.4.
doi: 10.1002/0471142905.hg1904s63.

Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using blue native gel electrophoresis

Francisca Díaz  1 , Antoni Barrientos, Flavia Fontanesi
Affiliations

VSports app下载 - Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using blue native gel electrophoresis

Francisca Díaz 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 progressively developed VSports手机版. Based on methods described elsewhere, the use of blue native gel electrophoresis (BNGE) techniques to routinely assess the OXPHOS system and screen for enzymatic defects in homogenates or mitochondrial preparations from tissues or cultured cells is described here. .

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Figures

Figure 1
Figure 1
Blue Native Gel electrophoresis. (A) Mouse muscle mitochondria separated by BNG and stained with Coomassie blue to visualize respiratory complexes, lane 1: control mouse, lane 2: COX deficient mouse, lane 3: Native High Molecular weight marker (GE). (B) BN-western blot using Invitrogen 4–16 gradient gel and Antibody Cocktail from Mitosciences. The samples are mitochondrial fractions from mouse fibroblasts. Lanes 1 and 3 control cells and lanes 2 and 4 fibroblasts deficient in CI and CIII. (C) In gel activity stain of respiratory complexes of mouse liver mitochondria. In CV activity stain the F1 (lower band) dissociates from the fully assembled complex (arrow) and retains ATP hydrolysis activity.
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References

    1. Acin-Perez R, Fernandez-Silva P, Peleato ML, Perez-Martos A, Enriquez JA. Respiratory active mitochondrial supercomplexes. Mol. Cell. 2008;32:529–539. - PubMed
    1. Calvaruso MA, Smeitink J, Nijtmans L. Electrophoresis techniques to investigate defects in oxidative phosphorylation. Methods. 2008;46:281–287. - PubMed
    1. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–685. - PubMed (V体育安卓版)
    1. Nijtmans LG, Henderson NS, Holt IJ. Blue Native electrophoresis to study mitochondrial and other protein complexes. Methods. 2002;26:327–334. - PubMed
    1. Schagger H, von Jagow G. Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal. Biochem. 1991;199:223–231. - PubMed

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