McDowell L M, Schmidt A, Cohen E R, Studelska D R, Schaefer J
Department of Chemistry Washington University, St. Louis, MO 63130, USA.
J Mol Biol. 1996 Feb 16;256(1):160-71. doi: 10.1006/jmbi.1996.0074.
The 46 kDa enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase catalyzes the condensation of shikimate-3-phosphate (S3P) and phosphoenolpyruvate to form EPSP. The reaction is inhibited by N-(phosphonomethyl)-glycine (Glp), which in the presence of S3P, binds to EPSP synthase to form a stable ternary complex. As part of a solid-state NMR characterization of this structure, 15N labels were introduced selectively into the lysine, arginine and histidine residues of EPSP synthase and distances to a 13C label in Glp and to the 31P in S3P and Glp were measured by rotational-echo double-resonance NMR. Three lysine and four arginine residues are in the proximity of the phosphate group of S3P and the carboxyl and phosphonate groups of Glp. A single histidine residue is in the vicinity of the binding site (closer to Glp than to S3P) but is more distant than the lysine and arginine residues.
46 kDa的5-烯醇丙酮酸莽草酸-3-磷酸（EPSP）合酶催化莽草酸-3-磷酸（S3P）与磷酸烯醇丙酮酸缩合形成EPSP。该反应受到N-（膦酰基甲基）-甘氨酸（Glp）的抑制，在S3P存在的情况下，Glp与EPSP合酶结合形成稳定的三元复合物。作为对该结构进行固态核磁共振表征的一部分，将15N标记选择性地引入到EPSP合酶的赖氨酸、精氨酸和组氨酸残基中，并通过旋转回波双共振核磁共振测量与Glp中的13C标记以及S3P和Glp中的31P的距离。三个赖氨酸残基和四个精氨酸残基靠近S3P的磷酸基团以及Glp的羧基和膦酸基团。单个组氨酸残基位于结合位点附近（比S3P更靠近Glp），但比赖氨酸和精氨酸残基距离更远。
