Binding site identification and structure determination of protein-ligand complexes by NMR a semiautomated approach.
Binding site identification and structure determination of protein-ligand complexes by NMR a semiautomated approach.
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通过NMR半荷兰方法对蛋白质配体复合物的结合位点鉴定和结构测定。
DOI：
10.1016/b978-0-12-381274-2.00010-8
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发表时间：
2011
影响因子：
--
通讯作者：
Volkman, Brian F.
中科院分区：
文献类型：
--
作者：
Ziarek, Joshua J.;Peterson, Francis C.;Lytle, Betsy L.;Volkman, Brian F.
在过去的十五年中，NMR光谱在药物发现的先导物鉴定和优化阶段的作用稳步增加。NMR在药物样化合物的生物物理分析中占据独特的位置，因为它能够在单个氨基酸的水平上识别结合位点，亲和力和配体姿势，而不必解决蛋白质-配体复合物的结构。然而，它也可以提供柔性蛋白质和低亲和力（Kd > 10-6 M）复合物的结构，这些复合物通常不能结晶。本文强调了一个以吞吐量为重点的协议，旨在确定结合位点的表征，自动化和半自动化的NMR分配方法，并通过NMR确定蛋白质-配体复合物的结构的实际方面。
Over the last fifteen years, the role of NMR spectroscopy in the lead identification and optimization stages of pharmaceutical drug discovery has steadily increased. NMR occupies a unique niche in the biophysical analysis of drug-like compounds because of its ability to identify binding sites, affinities, and ligand poses at the level of individual amino acids without necessarily solving the structure of the protein-ligand complex. However, it can also provide structures of flexible proteins and low-affinity (Kd > 10-6 M) complexes, which often fail to crystallize. This article emphasizes a throughput-focused protocol that aims to identify practical aspects of binding site characterization, automated and semi-automated NMR assignment methods, and structure determination of protein-ligand complexes by NMR.