Wang Yanli, Liu Lin, Wei Zhiyi, Cheng Zhongjun, Lin Yajing, Gong Weimin
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
J Biol Chem. 2006 Dec 22;281(51):39642-8. doi: 10.1074/jbc.M606421200. Epub 2006 Oct 18.
Bisphosphoglycerate mutase is an erythrocyte-specific enzyme catalyzing a series of intermolecular phosphoryl group transfer reactions. Its main function is to synthesize 2,3-bisphosphoglycerate, the allosteric effector of hemoglobin. In this paper, we directly observed real-time motion of the enzyme active site and the substrate during phosphoryl transfer. A series of high resolution crystal structures of human bisphosphoglycerate mutase co-crystallized with 2,3-bisphosphoglycerate, representing different time points in the phosphoryl transfer reaction, were solved. These structures not only clarify the argument concerning the substrate binding mode for this enzyme family but also depict the entire process of the key histidine phosphorylation as a "slow movie". It was observed that the enzyme conformation continuously changed during the different states of the reaction. These results provide direct evidence for an "in line" phosphoryl transfer mechanism, and the roles of some key residues in the phosphoryl transfer process are identified.
二磷酸甘油酸变位酶是一种红细胞特异性酶，催化一系列分子间磷酰基转移反应。其主要功能是合成2,3-二磷酸甘油酸，即血红蛋白的变构效应剂。在本文中，我们直接观察到了磷酰基转移过程中酶活性位点和底物的实时运动。解析了一系列与2,3-二磷酸甘油酸共结晶的人二磷酸甘油酸变位酶的高分辨率晶体结构，这些结构代表了磷酰基转移反应中的不同时间点。这些结构不仅阐明了关于该酶家族底物结合模式的争论，还将关键组氨酸磷酸化的整个过程描绘成一部“慢动作影片”。据观察，在反应的不同状态下，酶的构象不断变化。这些结果为“线性”磷酰基转移机制提供了直接证据，并确定了磷酰基转移过程中一些关键残基的作用。
