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Kinetic analysis of lactate dehydrogenase using integrated rate equations
L D Holmes1, M R Schiller, E A Boeker
1Department of Chemistry and Biochemistry, Utah State University, Logan 84322.
Experientia
|October 15, 1993
Summary
Lactate dehydrogenase kinetics were analyzed using integrated rate equations, revealing a ternary complex mechanism. This study precisely determined kinetic constants and oxamate inhibition, supporting the proposed reaction pathway.
Area of Science:
- Biochemistry
- Enzyme kinetics
Background:
- Lactate dehydrogenase (LDH) is crucial in cellular metabolism.
- Understanding LDH reaction mechanisms is vital for biochemical research.
Purpose of the Study:
- To analyze the lactate dehydrogenase reaction mechanism under second-order conditions.
- To determine kinetic constants and substrate inhibition parameters.
- To validate the ternary complex mechanism for LDH.
Main Methods:
- Utilized integrated rate equations for kinetic analysis.
- Employed a two-step regression analysis on sextuplicate progress curves.
- Applied a second-order integrated rate equation with substrate inhibition terms.
Main Results:
- Achieved a fitting error of less than one percent.
- Determined kinetic constants exclusively supporting a ternary complex mechanism.
- Quantified the inhibition constant for oxamate.
Conclusions:
- The kinetic data strongly support a ternary complex mechanism for lactate dehydrogenase.
- The study provides precise kinetic parameters for LDH.
- Oxamate was confirmed as an inhibitor with a determined inhibition constant.