Book Chapter

Assessment of mitochondrial respiratory chain enzymes in cells and tissues

Ann E Frazier, Amy E Vincent, Doug M Turnbull, David R Thorburn, Robert W Taylor

MITOCHONDRIA, 3RD EDITION | Methods in Cell Biology | ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD | Published : 2020

Abstract

Measurement of the individual enzymes involved in mitochondrial oxidative phosphorylation (OXPHOS) forms a key part of diagnostic investigations in patients with suspected mitochondrial disease, and can provide crucial information on mitochondrial OXPHOS function in a variety of cells and tissues that are applicable to many research investigations. In this chapter, we present methods for analysis of mitochondrial respiratory chain enzymes in cells and tissues based on assays performed in two geographically separate diagnostic referral centers, as part of clinical diagnostic investigations. Techniques for sample preparation from cells and tissues, and spectrophotometric assays for measurement..

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Grants

Awarded by Wellcome Trust Centre for Mitochondrial Research


Awarded by Mitochondrial Disease Patient Cohort (UK)


Awarded by Newcastle University Centre for Ageing and Vitality (Medical Research Council)


Awarded by Sir Henry Wellcome Fellowship


Awarded by Australian National Health & Medical Research Council (NHMRC)


Funding Acknowledgements

The authors would like to thank Simone Tregoning and Tegan Stait for assistance with the respiratory chain enzymology and Alison Compton and Charlotte Warren for contribution of panels to Figs. 3 and 4, respectively. R.W.T. and D.M.T. are supported by the Wellcome Trust Centre for Mitochondrial Research (203105/Z/16/Z), the Medical Research Council (MRC) International Centre for Genomic Medicine in Neuromuscular Disease, the Mitochondrial Disease Patient Cohort (UK) (G0800674), Newcastle University Centre for Ageing and Vitality (supported by the Biotechnology and Biological Sciences Research Council and Medical Research Council L016354), UK NIHR Biomedical Research Centre for Ageing and Agerelated disease award to the Newcastle upon Tyne Hospitals NHS Foundation Trust, the Lily Foundation and the UK NHS Specialist Commissioners which funds the "Rare Mitochondrial Disorders of Adults and Children" Diagnostic Service in Newcastle upon Tyne (http://www.newcastle-mitochondria.com/).A.E.V.is the recipient of a Sir Henry Wellcome Fellowship (215888/Z/19/Z). D.R.T. is supported by grants (GNT1164479, GNT1107094, GNT1113531) and a Principal Research Fellowship (GNT1155244) from the Australian National Health & Medical Research Council (NHMRC), by a Vincent Chiodo Charitable Trust Fellowship, along with funding to A.E.F. and D.R.T. from the Mito Foundation and by the Victorian Government's Operational Infrastructure Support Program.