De la Mare S, Coulson A F, Knowles J R, Priddle J D, Offord R E
Biochem J. 1972 Sep;129(2):321-31. doi: 10.1042/bj1290321.
Triose phosphate isomerase from chicken muscle reacts stoicheiometrically with the active-site-directed irreversible inhibitor bromohydroxyacetone phosphate with concomitant loss of all catalytic activity. The primary site of attachment has been shown to be a unique glutamic acid residue in the sequence Ala-Tyr-Glu-Pro-Val-Trp. Unless the inhibitor-enzyme bond is stabilized by reduction of the C-2 carbonyl group with borohydride, the phosphate group is lost and the label migrates to the adjacent tyrosine residue. It is suggested that the gamma-carboxylate group of the glutamic acid residue may be the base responsible for primary proton abstraction from substrate in the catalysis. The failure of this reagent specifically to inactivate either muscle or yeast aldolase, and the use of the reagent in preparing isomerase-free glycolytic enzymes, is discussed.
来自鸡肌肉的磷酸丙糖异构酶与活性位点导向的不可逆抑制剂磷酸溴代羟基丙酮发生化学计量反应，同时所有催化活性丧失。已证明结合的主要位点是序列Ala-Tyr-Glu-Pro-Val-Trp中一个独特的谷氨酸残基。除非通过硼氢化物还原C-2羰基使抑制剂-酶键稳定，否则磷酸基团会丢失，标记会迁移到相邻的酪氨酸残基上。有人提出，谷氨酸残基的γ-羧基可能是催化过程中从底物上夺取初级质子的碱。讨论了该试剂不能特异性灭活肌肉或酵母醛缩酶的情况，以及该试剂在制备无异构酶糖酵解酶中的应用。
