Refinement of the conformation of UDP-galactose bound to galactosyltransferase using the STD NMR intensity-restrained CORCEMA optimization.
Refinement of the conformation of UDP-galactose bound to galactosyltransferase using the STD NMR intensity-restrained CORCEMA optimization.
复制标题
使用 STD NMR 强度限制 CORCEMA 优化对与半乳糖基转移酶结合的 UDP-半乳糖的构象进行细化。
DOI：
10.1021/ja048703u
复制
发表时间：
2004
影响因子：
15
通讯作者：
Krishna,NRama
中科院分区：
文献类型：
--
作者：
Jayalakshmi,V;Biet,Thorsten;Peters,Thomas;Krishna,NRama
STD核磁共振技术最初被描述为一种筛选大型化合物文库的工具，以确定对感兴趣的目标蛋白质具有特异性的先导化合物。还描述了该技术在蛋白质、病毒衣壳和核酸弱结合配体的定性表位定位中的应用。在这里，我们描述了应用标准核磁共振强度受限的CORCEMA优化(SICO)程序来精制半乳糖转移酶复合体中UDP−半乳糖的结合构象，以500 MHz记录的标准核磁共振强度作为实验约束。将结合的UdP−半乳糖在溶液中的SiCO结构与该络合物晶体结构中的SICO结构进行比较，发现该蛋白质在配体扭转角和V253侧链方向上有所不同。这项工作描述了第一次应用STD核磁共振强度受限的CORCEMA优化程序来精炼结合配体结构的扭转角。这种方法很可能在基于结构的药物设计程序中有用，因为大多数初始的先导化合物通常对药物感兴趣的目标蛋白表现出弱的亲和力(毫摩尔到微摩尔)，并且这些先导化合物在蛋白质结合口袋中的结合构象可以通过CORCEMA-ST精化来确定。
The STD NMR technique has originally been described as a tool for screening large compound libraries to identify the lead compounds that are specific to target proteins of interest. The application of this technique in the qualitative epitope mapping of ligands weakly binding to proteins, virus capsid shells, and nucleic acids has also been described. Here we describe the application of the STD NMR intensity-restrained CORCEMA optimization (SICO) procedure for refining the bound conformation of UDP−galactose in galactosyltransferase complex using STD NMR intensities recorded at 500 MHz as the experimental constraints. A comparison of the SICO structure for the bound UDP−galactose in solution with that in the crystal structure for this complex shows some differences in ligand torsion angles and V253 side-chain orientation in the protein. This work describes the first application of an STD NMR intensity-restrained CORCEMA optimization procedure for refining the torsion angles of a bound ligand structure. This method is likely to be useful in structure-based drug design programs since most initial lead compounds generally exhibit weak affinity (millimolar to micromolar) to target proteins of pharmaceutical interest, and the bound conformation of these lead compounds in the protein binding pocket can be determined by the CORCEMA-ST refinement.