The determination and analysis of site-specific rates of mitochondrial reactive oxygen species production

Methods Enzymol. 2013;526:189-217. doi: 10.1016/B978-0-12-405883-5.00012-0.


Mitochondrial reactive oxygen species (ROS) are widely implicated in physiological and pathological pathways. We propose that it is critical to understand the specific sites of mitochondrial ROS production and their mechanisms of action. Mitochondria possess at least eight distinct sites of ROS production in the electron transport chain and matrix compartment. In this chapter, we describe the nature of the mitochondrial ROS-producing machinery and the relative capacities of each site. We provide detailed methods for the measurement of H2O2 release and the conditions under which maximal rates from each site can be achieved in intact skeletal muscle mitochondria.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biochemistry / methods
  • Electron Transport Complex I / metabolism
  • Electron Transport Complex II / metabolism
  • Electron Transport Complex III / metabolism
  • Glycerolphosphate Dehydrogenase / metabolism
  • Humans
  • Hydrogen Peroxide / analysis*
  • Hydrogen Peroxide / metabolism
  • Mitochondria, Muscle / metabolism*
  • Muscle, Skeletal / metabolism
  • Reactive Oxygen Species / analysis*
  • Reactive Oxygen Species / metabolism


  • Reactive Oxygen Species
  • Hydrogen Peroxide
  • Glycerolphosphate Dehydrogenase
  • Electron Transport Complex II
  • Electron Transport Complex I
  • Electron Transport Complex III