Measurements of ATP in Mammalian Cells

Methods. 2002 Apr;26(4):317-26. doi: 10.1016/S1046-2023(02)00037-3.

Abstract

Levels of phosphorylated adenosine nucleotides, including the universal energy carrier adenosine 5(')-triphosphate (ATP) and its metabolites adenosine 5(')-diphosphate (ADP) and adenosine 5(')-monophosphate (AMP), define the energy state in living cells and are dependent mainly on mitochondrial function. In this article, we describe a method based on the luciferase-luciferin system used to measure mitochondrial ATP synthesis continuously in permeabilized mammalian cells and mitochondria isolated from animal tissues. We also describe a technique that uses the expression of recombinant targeted luciferase to report ATP content in different cell compartments. Finally, we describe an HPLC-based method for accurate measurement of ATP, ADP, and AMP in cultured cells and animal tissues.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Diphosphate / metabolism
  • Adenosine Monophosphate / metabolism
  • Adenosine Triphosphate / analysis*
  • Adenosine Triphosphate / metabolism
  • Animals
  • Biochemistry / methods*
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Creatine / metabolism
  • Firefly Luciferin / metabolism
  • Kinetics
  • Luciferases / metabolism
  • Mice
  • Mitochondria / metabolism*
  • Mitochondria, Liver / metabolism
  • Models, Chemical
  • Plasmids / metabolism
  • Rats
  • Time Factors
  • Tumor Cells, Cultured

Substances

  • Adenosine Monophosphate
  • Firefly Luciferin
  • Adenosine Diphosphate
  • Adenosine Triphosphate
  • Luciferases
  • Creatine