Measurements of ATP in mammalian cells.
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.
Manfredi
Giovanni
G
Department of Neurology and Neuroscience, Weill Medical College of Cornell University, 525 East 68th Street, A-505, New York, USA. gim2004@mail.med.cornell.edu
Yang
Lichuan
L
Gajewski
Carl D
CD
Mattiazzi
Marina
M
D000244:Adenosine Diphosphate / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003401:Creatine / Q000378:metabolism; D049454:Firefly Luciferin / Q000378:metabolism; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D008956:Models, Chemical; D010957:Plasmids / Q000378:metabolism; D051381:Rats; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000249:Adenosine Monophosphate; D049454:Firefly Luciferin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D008156:Luciferases; D003401:Creatine