Molecular Pharmacology, Vol 6, 31-40, Copyright © 1970 by the American Society for Pharmacology and Experimental Therapeutics

A Kinetic Model of a Cyclic System for the Fluorometric Microdetermination of Adenosine Triphosphatase Activity

¹ Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland 21201

The derivation of a series of kinetic rate equations to model a cyclic, multistep, fluorometric assay for ADP-generating systems is described. The reaction curves predicted by the model equations are compared with experimentally observed velocities for ATPase reactions to show that reaction rates that are constant and rate-limited by the rate of ADP generation are easily obtained within practical limits of auxiliary enzyme concentrations. The potential of this technique for the study of substrate-enzyme interactions over wide ranges of substrate and enzyme concentrations is discussed.

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ACKNOWLEDGMENTS The author wishes to express his gratitude to Dr. John J. O'Neill and Dr. William D. Blake for their invaluable advice and encouragement.