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Biochemical Analyses of the Electron Transport Chain Complexes by Spectrophotometry

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Mitochondrial Disorders

Part of the book series: Methods in Molecular Biology ((MIMB,volume 837))

Abstract

In the diagnostic work-up of patients with suspected mitochondrial disease, evaluating the activity of the individual oxidative phosphorylation (OXPHOS) complexes is crucial. Here, we describe spectrophotometric assays for OXPHOS enzymology that can be applied to both tissue samples and cultured cells. These assays are designed to assess the enzymatic activity of the individual OXPHOS complexes I–V, along with the Krebs cycle enzyme citrate synthase as a mitochondrial control. As well, we include an assay for the coupled energy transfer between complexes II and III. Determining the enzymatic activities can be valuable in defining isolated or multicomplex disorders and may be relevant to the design of future molecular investigations.

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Acknowledgements

Special thanks to Simone Tregoning and Wendy Salter for assistance with diagnostic data. This work was supported by grants from the Australian National Health and Medical Research Council (NHMRC) and the Muscular Dystrophy Foundation, an NHMRC Principal Research Fellowship to David Thorburn, and an NHMRC Career Development Award to Ann Frazier.

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Authors and Affiliations

  1. Murdoch Children’s Research Institute, Parkville, VIC, Australia

    Ann E. Frazier

  2. Murdoch Childrens Research Institute and Victorian Clinical Genetics Services Pathology, Royal Children’s Hospital, Melbourne, VIC, Australia

    David R. Thorburn

  3. Department of Paediatrics, University of Melbourne, Melbourne, VIC, Australia

    David R. Thorburn

Authors
  1. Ann E. Frazier
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  2. David R. Thorburn
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Corresponding author

Correspondence to David R. Thorburn .

Editor information

Editors and Affiliations

  1. , Molecular and Human Genetics, Baylor College of Medicine, 1 Baylor Plaza, Houston, 77030, Texas, USA

    Lee-Jun C. Wong, Ph.D.

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Frazier, A.E., Thorburn, D.R. (2012). Biochemical Analyses of the Electron Transport Chain Complexes by Spectrophotometry. In: Wong, Ph.D., LJ. (eds) Mitochondrial Disorders. Methods in Molecular Biology, vol 837. Humana Press. https://doi.org/10.1007/978-1-61779-504-6_4

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  • DOI: https://doi.org/10.1007/978-1-61779-504-6_4

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-503-9

  • Online ISBN: 978-1-61779-504-6

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