Transient-state kinetic approach to mechanisms of enzymatic catalysis.
Transient-state kinetics by its inherent nature can potentially provide more directly observed detailed resolution of discrete events in the mechanistic time courses of enzyme-catalyzed reactions than its more widely used steady-state counterpart. The use of the transient-state approach, however, has been severely limited by the lack of any theoretically sound and applicable basis of interpreting the virtual cornucopia of time and signal-dependent phenomena that it provides. This Account describes the basic kinetic behavior of the transient state, critically examines some currently used analytic methods, discusses the application of a new and more soundly based "resolved component transient-state time-course method" to the L-glutamate-dehydrogenase reaction, and establishes new approaches for the analysis of both single- and multiple-step substituted transient-state kinetic isotope effects.
Fisher
Harvey F
HF
Laboratory of Molecular Biochemistry, Veterans Affairs Medical Center, Department of Biochemistry, University of Kansas Medical Center, 4801 Linwood Boulevard, Kansas City, MO 64128-2226, USA. hfisher@kumc.edu
D002384:Catalysis; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism*; D005969:Glutamate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008433:Mathematics; D015394:Molecular Structure; D011487:Protein Conformation
D004798:Enzymes; D005969:Glutamate Dehydrogenase