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Use of a Real-Time, Coupled Assay to Measure the ATPase Activity of DNA Topoisomerase II

  • Janet E. Lindsley
Part of the Methods in Molecular Biology™ book series (MIMB, volume 95)

Abstract

This chapter describes the use of a common spectrophotometric assay for following the rate of ATP hydrolysis as applied to type II DNA topoisomerases. It is called a “coupled assay,” because each time an ATP molecule is hydro-lyzed, a molecule of NADH is rapidly oxidized; ATP hydrolysis and NADH oxidation are therefore coupled. Although NADH absorbs strongly at 340 nm, the product of its oxidation, NAD+, does not. Therefore, the rate of ATP hydrolysis can be determined by following the decrease in optical absorbance of the reaction at 340 nm.

Keywords

ATPase Activity Lactate Dehydrogenase Pyruvate Kinase NADH Oxidation Coupling Enzyme 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Janet E. Lindsley

There are no affiliations available

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