Kinetic Analysis of Macromolecules: A Practical Approach (Practical Approach Series)

Modern enzymology can no longer rely upon indirect inferences derived by steady state kinetic analysis because these are often misleading. Rather, in the era of 'New Enzymology', reactions at the active sites of enzyme are monitored by direct measurement of individual steps in the sequence to establish the pathway of reactions occurring. These quantitative methods can be directly related to high resolution structural information and define the underlying kinetic and thermodynamic basis for catalysis. The goal of a comprehensive kinetic analysis is to identify each step in the reaction pathway, define the rate and equilibrium constants for each step, identify all conformational and chemical intermediates, identify the rate-limiting steps, and ultimately establish the basis for enzymatic specificity and efficiency. This volume contains chapters contributed by the leading experts in the application of modern kinetic methods to complex problems in the study of catalysis by both proteins and RNA catalysts. Each chapter focuses on a unique system and the novel methods used to obtain definitive data that can be directly applied to structural information, giving the much-needed practical details that are often omitted in the general literature. Readers will benefit greatly from the richness of each chapter in describing what is most important in solving the most difficult problems in modern enzymology.