Measurements of ATP in mammalian cells

被引:213
作者
Manfredi, G [1 ]
Yang, LC [1 ]
Gajewski, CD [1 ]
Mattiazzi, M [1 ]
机构
[1] Cornell Univ, Weill Med Coll, Dept Neurol & Neurosci, New York, NY 10021 USA
关键词
ATP; mitochondria; ATP synthesis; high-performance liquid chromatography; luciferase; luciferin;
D O I
10.1016/S1046-2023(02)00037-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2002 Elsevier Science (USA). All rights reserved.
引用
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页码:317 / 326
页数:10
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