Contemporary Enzyme Kinetics and Mechanism: Selected Methods in Enzymology

Kinetic studies of enzyme action provide powerful insights into the underlying mechanisms of catalysis and regulation. These approaches are equally useful in examining the action of newly discovered enzymes and therapeutic agents.Contemporary Enzyme Kinetics and Mechanism, Second Edition presents key articles from Volumes 63, 64, 87, and 249 of Methods in Enzymology. The chapters describe the most essential and widely applied strategies. A set of exercises and problems is included to facilitate mastery of these topics.The book will aid the reader to design, execute, and analyze kinetic experiments on enzymes. Its emphasis on enzyme inhibition will also make it attractive to pharmacologists and pharmaceutical chemists interested in rational drug design.Of the seventeen chapters presented in this new edition, ten did not previously appear in the first edition. Transient kinetic approaches to enzyme mechanisms Designing initial rate enzyme assay Deriving initial velocity and isotope exchange rate equations Plotting and statistical methods for analyzing rate data Cooperativity in enzyme function Reversible enzyme inhibitors as mechanistic probes Transition-state and multisubstrate inhibitors Affinity labeling to probe enzyme structure and function Mechanism-based enzyme inactivators Isotope exchange methods for elucidating enzymatic catalysis Kinetic isotope effects in enzyme catalysis Site-directed mutagenesis in studies of enzyme catalysis