Oxidative phosphorylation measurement in cell lines and tissues

Mitochondrial oxidative phosphorylation (OXPHOS) dysfunction is implicated in a growing spectrum of diseases, from neurodegeneration to cancer. Where tissues or transformed cells are available, respirometry and enzymology allow a sophisticated analysis of OXPHOS with modest-cost equipment. The isola...

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Bibliographic Details
Published in:Mitochondrion 2014-03, Vol.15, p.24-33
Main Authors: Van Bergen, Nicole J, Blake, Rachel E, Crowston, Jonathan G, Trounce, Ian A
Format: Article
Language:English
Subjects:
Cell Line
Cytological Techniques - methods
Humans
Mitochondria - metabolism
Oxidative Phosphorylation
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Summary:Mitochondrial oxidative phosphorylation (OXPHOS) dysfunction is implicated in a growing spectrum of diseases, from neurodegeneration to cancer. Where tissues or transformed cells are available, respirometry and enzymology allow a sophisticated analysis of OXPHOS with modest-cost equipment. The isolation of organelle fractions is also invaluable for determining association of proteins of interest. Here we revisit and consolidate methods to measure whole cell mitochondrial ATP synthesis, respiration, isolation of mitochondria from cultured cells and tissues, and OXPHOS enzymology. We also explain common pitfalls, guide optimisation of the methods for new users, and provide full laboratory protocols in Supplementary materials.
ISSN:1567-7249
1872-8278
DOI:10.1016/j.mito.2014.03.003