SOS-NMR: A saturation transfer NMR-based method for determining the structures of protein-ligand complexes
SOS-NMR: A saturation transfer NMR-based method for determining the structures of protein-ligand complexes
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DOI：
10.1021/ja039480v
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发表时间：
2004-03-03
影响因子：
15
通讯作者：
Beutel, BA
中科院分区：
文献类型：
--
作者：
Hajduk, PJ;Mack, JC;Beutel, BA
一个基于NMR的替代传统的X-射线晶体学和NMR方法的结构为基础的药物设计的描述，使配体的结构确定复杂的几乎任何生物分子的目标，无论大小，组成，或低聚状态。该方法利用饱和转移差（STD）NMR光谱上进行的配体络合到一系列的目标样品已被氘化的地方，除了特定的氨基酸类型。以这种方式，可以定义配体结合位点的氨基酸组成，并且，给定蛋白质靶标的三维结构，可以确定蛋白质-配体复合物的三维结构。不像早期的NMR方法解决蛋白质-配体复合物的结构，没有蛋白质共振分配是必要的。因此，该方法具有广泛的潜在应用，特别是在X射线晶体学和传统NMR方法无法产生结构数据的情况下。该方法被称为SOS-NMR，使用Overhauser效应和选择性标记获得结构信息，并在两种蛋白质-配体复合物上进行了验证：与2-（3 '-吡啶基）-苯并咪唑复合的FKBP和与尿苷二磷酸N-乙酰葡糖胺复合的MurA。
An NMR-based alternative to traditional X-ray crystallography and NMR methods for structure-based drug design is described that enables the structure determination of ligands complexed to virtually any biomolecular target regardless of size, composition, or oligomeric state. The method utilizes saturation transfer difference (STD) NMR spectroscopy performed on a ligand complexed to a series of target samples that have been deuterated everywhere except for specific amino acid types. In this way, the amino acid composition of the ligand-binding site can be defined, and, given the three-dimensional structure of the protein target, the three-dimensional structure of the protein-ligand complex can be determined. Unlike earlier NMR methods for solving the structures of protein-ligand complexes, no protein resonance assignments are necessary. Thus, the approach has broad potential applications-especially in cases where X-ray crystallography and traditional NMR methods have failed to produce structural data. The method is called SOS-NMR for structural information using Overhauser effects and selective labeling and is validated on two protein-ligand complexes: FKBP complexed to 2-(3'-pyridyl)-benzimidazole and MurA complexed to uridine diphosphate N-acetylglucosamine.