Time-resolved solid-state REDOR NMR studies of UDP N-acetylglucosamine enolpyruvyl transferase.
Time-resolved solid-state REDOR NMR studies of UDP N-acetylglucosamine enolpyruvyl transferase.
复制标题
UDP N-乙酰氨基葡萄糖烯醇丙酮基转移酶的时间分辨固态 REDOR NMR 研究。
DOI：
10.1016/0014-5793(95)01338-5
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发表时间：
1995
期刊：
影响因子：
3.5
通讯作者：
Evans,JN
中科院分区：
文献类型：
--
作者：
Li,Y;Krekel,F;Ramilo,CA;Amrhein,N;Evans,JN
本实验室最近推出的时间分辨固体旋转回波双共振(REDOR)核磁共振波谱新方法，应用于尿苷N-乙酰氨基葡萄糖(UDP-NAG)烯醇式丙酮基转移酶(EPT)，目的是探索活性物种与其酶活性部位之间的相互作用。该方法已被定性地用于15N标记的UDP-NAG EPT的酶抑制剂和酶中间体复合体，在稳态和预稳态条件下被捕获。在时间分辨条件下(反应时间50ms后)，底物UDP-NAG、UDP-NAG+3-Z-磷烯醇式丙酮酸、共价O-磷酸硫酮和UDP-NAG+磷烯醇式丙酮酸与EPT酶活性部位之间发生了一组不同的分子间相互作用，这与相关酶5-烯醇式丙酮酸-莽草酸-3-磷酸合成酶的相互作用的类似研究形成了对比。
The new method of time-resolved solid-state rotational echo double resonance (REDOR) NMR spectroscopy introduced recently by this laboratory has been applied to the enzyme uridine N-acetylglucosamine (UDP-NAG) enolpyruvyl transferase (EPT), with the goal of probing the interactions between reactive species and their enzyme active site. The approach has been used in a qualitative fashion with the enzyme-inhibitor and enzyme-intermediate complexes of uniformly15N-labeled UDP-NAG EPT, trapped under steady-state and pre-steady-state conditions. A different set of intermolecular interactions between the substrates UDP-NAG, UDP-NAG plus 3-Z-fluorophosphoenolpyruvate, covalent O-phosphothioketal, and UDP-NAG plus phosphoenolpyruvate trapped under time-resolved conditions (after 50 ms reaction time), and the EPT enzyme active site were observed, and this is contrasted to a similar study of the interactions in a related enzyme, 5-enolpyruvyl-shikimate-3-phosphate synthase.