Use of selective Trp side chain labeling to characterize protein-protein and protein-ligand interactions by NMR spectroscopy.
Use of selective Trp side chain labeling to characterize protein-protein and protein-ligand interactions by NMR spectroscopy.
复制标题
使用选择性色氨酸侧链标记通过核磁共振波谱表征蛋白质-蛋白质和蛋白质-配体相互作用。
DOI：
--
复制
发表时间：
2003
影响因子：
15
通讯作者：
M. Pellecchia
中科院分区：
文献类型：
--
作者：
R. Rodríguez;M. Pellecchia
最近的研究表明，在蛋白质结合位点的氨基酸发生只有减少数量的残基负责蛋白质-蛋白质和蛋白质-配体相互作用的大部分相互作用能。总之，色氨酸（Trp）似乎是蛋白质热点中最常见的残基。在这里，我们报告了一种新的，有效的，具有成本效益的方法，选择性地将特定的同位素标记到重组蛋白的色氨酸残基的侧链。我们表明，所提出的方法允许选择性NMR观察色氨酸侧链，使配体结合，蛋白质-蛋白质相互作用，氢键结合，蛋白质折叠，侧链动力学的研究。将给出蛋白质BIR 3的实例。
Recent studies on amino acid occurrence in protein binding sites suggest that only a reduced number of residues are responsible for most interaction energy in protein-protein and protein-ligand interactions. Above all, tryptophan (Trp) seems to be the most frequent residue in protein's hot spots. Here we report a novel, efficient, and cost-effective method to selectively incorporate specific isotope labels into the side chains of Trp residues in recombinant proteins. We show that the method proposed allows selective NMR observation of Trp side chains that enables studies of ligand binding, protein-protein interactions, hydrogen binding, protein folding, and side chain dynamics. Examples with the protein BIR3 will be given.