The Department of Biochemistry, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, Kansas 66160, USA.
Biochemistry. 2009 Dec 29;48(51):12265-71. doi: 10.1021/bi901514h.
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.
瞬态动力学方法能够实时揭示中间产物的形成和后续转化。然而，与稳态方法中基于代数算法的间接但可靠的理论基础相比，瞬态动力学方法缺乏严格适用的理论基础，这极大地限制了其在酶促反应和其他复杂化学反应机制的机理解析方面的应用。最近，我们建立了三个严格推导的瞬态动力学基本“规则”，适用于实际的多步反应，在此，我们成功地将非常违反直觉的“第二规则”应用于解析 l-苯丙氨酸脱氢酶催化反应的机制。
