Biochemical analyses of the electron transport chain complexes by spectrophotometry

Methods Mol Biol. 2012;837:49-62. doi: 10.1007/978-1-61779-504-6_4.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Citrate (si)-Synthase / metabolism
  • Electron Transport Chain Complex Proteins / metabolism*
  • Enzyme Assays / methods*
  • Humans
  • Mitochondria / enzymology
  • Mitochondria / metabolism
  • Oxidative Phosphorylation
  • Spectrophotometry / methods*


  • Electron Transport Chain Complex Proteins
  • Citrate (si)-Synthase