Structural constraints on the ternary complex of 5-enolpyruvylshikimate-3-phosphate synthase from rotational-echo double-resonance NMR
Structural constraints on the ternary complex of 5-enolpyruvylshikimate-3-phosphate synthase from rotational-echo double-resonance NMR
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DOI：
10.1006/jmbi.1996.0074
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发表时间：
1996-02-16
影响因子：
5.6
通讯作者：
Schaefer, J
中科院分区：
文献类型：
--
作者：
McDowell, LM;Schmidt, A;Schaefer, J
4 6 kDa的5-烯醇式丙酮酸-3-磷酸(EPSP)合成酶催化莽草酸-3-磷酸(S3P)与磷酸烯醇式丙酮酸(S3P)缩合生成EPSP。N-(磷酸亚甲基)-甘氨酸(GLP)抑制该反应，在S3P存在下，GLP与EPSP合成酶结合形成稳定的三元络合物。作为这种结构的固体核磁共振表征的一部分，N-15标记被选择性地引入到EPSP合成酶的赖氨酸、精氨酸和组氨酸残基上，并通过旋转回波双共振核磁共振测量了GLP中C-13标记和S3P和GLP中P-31的距离。三个赖氨酸和四个精氨酸残基位于S3P的磷酸基团以及GLP的羧基和膦酸基附近。单个组氨酸残基位于结合位点附近(更接近GLP而不是S3P)，但比赖氨酸和精氨酸残基更远。(C)1996学术出版社有限公司。
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, N-15 labels were introduced selectively into the lysine, arginine and histidine residues of EPSP synthase and distances to a C-13 label in Glp and to the P-31 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. (C) 1996 Academic Press Limited.