Measurement of Proton Leak in Isolated Mitochondria

Methods Mol Biol. 2018:1782:157-170. doi: 10.1007/978-1-4939-7831-1_9.

Abstract

Oxidative phosphorylation is an important energy-conserving mechanism coupling mitochondrial electron transfer to ATP synthesis. Coupling between respiration and phosphorylation is not fully efficient due to proton leaks. In this chapter, we present a method to measure proton leak activity in isolated mitochondria. The relative strength of a modular kinetic approach to probe oxidative phosphorylation is emphasized.

Keywords: Membrane potential; Mitochondria; Modular kinetic analysis; Oxidative phosphorylation; Oxygen consumption.

MeSH terms

  • Adenosine Triphosphate / biosynthesis
  • Animals
  • Cell Respiration
  • Electrodes
  • Kinetics
  • Membrane Potential, Mitochondrial
  • Mitochondria / metabolism*
  • Mitochondrial Membranes / metabolism*
  • Muscle, Skeletal / cytology
  • Onium Compounds / metabolism
  • Oxidative Phosphorylation*
  • Oxygen / metabolism*
  • Oxygen Consumption
  • Protons*
  • Rats
  • Trityl Compounds / metabolism

Substances

  • Onium Compounds
  • Protons
  • Trityl Compounds
  • triphenylmethylphosphonium
  • Adenosine Triphosphate
  • Oxygen