Application of the second rule of transient-state kinetic isotope effects to an enzymatic mechanism.
The transient-state kinetic approach reveals the formation and subsequent interconversions of intermediates in real time. Its potential for the mechanistic resolution of enzymatic and other complex chemical mechanisms has been severely limited however by the lack of a rigorous and applicable theoretical basis in contrast to that of the less direct but soundly based algebraic algorithms of the steady-state approach. Having recently established three rigorously derived fundamental "rules" of transient-state kinetics applicable to realistic multiple step reactions, we present here the successful application of the very counterintuitive "second rule" to the resolution of the mechanism of the l-phenylalanine dehydrogenase catalyzed reaction.
Fisher
Harvey F
HF
The Department of Biochemistry, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, Kansas 66160, USA. hfisher@kumc.edu
Maniscalco
Steven J
SJ
Tally
Jon
J
Tabanor
Kayann
K
D000594:Amino Acid Oxidoreductases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D007554:Isotopes / Q000737:chemistry*; D007700:Kinetics; D012240:Rhodococcus / Q000737:chemistry / Q000201:enzymology*
D001426:Bacterial Proteins; D007554:Isotopes; D000594:Amino Acid Oxidoreductases; C022342:phenylalanine oxidase