Internal thermodynamics of enzymes determined by equilibrium quench: values of Kint for enolase and creatine kinase.
The equilibrium constant (Kint) for the enzyme-bound substrate and product of a one substrate/one product enzyme (enolase) and for those of a two substrate/two product enzyme (creatine kinase) have been determined. The values of Kint were determined by the rapid quenching of equilibrium mixtures of enzyme and radiolabeled substrate and product, under conditions where all of the marker substrate and product are bound. The scope and limitations of this method are discussed. Values of Kint have been collected from the literature, and it is shown that these data are consistent with the theory for kinetically optimized enzymes that is developed in the preceding paper.
Burbaum
J J
JJ
Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138.
Knowles
J R
JR
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D003402:Creatine Kinase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008954:Models, Biological; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D013379:Substrate Specificity; D013816:Thermodynamics
D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D003402:Creatine Kinase; D010751:Phosphopyruvate Hydratase