Determination of dissociation constants and specific rate constants of enzyme-substrate (or protein-ligand) interactions from rapid reaction kinetic data.
Determination of dissociation constants and specific rate constants of enzyme-substrate (or protein-ligand) interactions from rapid reaction kinetic data.
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根据快速反应动力学数据确定酶-底物（或蛋白质-配体）相互作用的解离常数和特定速率常数。
DOI：
10.1016/s0021-9258(19)41384-7
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
V. Massey
中科院分区：
文献类型：
--
作者：
S. Strickland;G. Palmer;V. Massey
近年来，能够在毫秒时间范围内跟踪吸光度或荧光变化的仪器越来越多，导致了大量的配体蛋白质相互作用的研究。酶学家特别感兴趣的是催化循环中可能发生部分反应的情况。例如，在有利的情况下，脱氢酶与还原的吡啶核苷酸的反应以及酶-辅酶复合物与参与催化反应的第二底物的反应之后可以是吸光度或荧光变化。古特弗罗因德最近的专着（1）讨论了许多这样的研究例子。同样适用于此类研究的是利用磷酸吡哆醛作为辅酶的酶，其中大的分光光度变化与反应机理中涉及的席夫碱中间体有关（2）。黄素蛋白具有广泛的光谱变化，与黄素的不同氧化还原状态以及酶-底物中间体有关，也构成了一个组，其中许多有用的信息已从停流研究中获得（例如，见参考文献3）。
The increasing availability of instruments capable of following absorbance or fluorescence changes in the millisecond time range has led in recent years to a large number of studies of ligandprotein interactions. Of particular interest to the enzymologist are cases where partial reactions in the catalytic cycle may be followed. For example, the reaction of a dehydrogenase with reduced pyridine nucleotide as well as the reaction of the enzymecoenzyme complex with the second substrate involved in the catalytic reaction may be followed in favorable cases by absorbance or fluorimetric changes. Numerous examples of such studies are discussed in the recent monograph by Gutfreund (1). Equally suited to such studies are enzymes utilizing pyridoxal phosphate as coenzyme, where large speetrophotometric changes are associated with the Schiff base intermediates involved in the reaction mechanisms (2). The flavoproteins, with extensive spectroscopic changes associated with the different oxidationreduction states of the flavin, as well as with enzyme-substrate intermediates, also constitute a group where much useful information has been obtained from stopped flow studies (see for example, Ref. 3).