Department of Molecular Genetics, The University of Toronto, ON, Canada.
J Biomol NMR. 2010 Jan;46(1):75-87. doi: 10.1007/s10858-009-9376-1. Epub 2009 Sep 27.
The development of new protein labeling strategies, along with optimized experiments that exploit the label, have significantly impacted on the types of biochemical problems that can now be addressed by solution NMR spectroscopy. Here we describe how methyl labeling of key residues in a highly deuterated protein background has facilitated studies of the structure, dynamics and interactions of supra-molecular particles. The methyl-labeling approach is briefly reviewed, followed by a summary of applications to three different molecular machines so as to illustrate the types of questions that can now be addressed. Areas where future innovations will lead to yet further improvements are highlighted as well.
新的蛋白质标记策略的发展,以及利用该标记优化实验,极大地影响了可以通过溶液 NMR 光谱学解决的生物化学问题的类型。在这里,我们描述了如何在高度氘化的蛋白质背景下对关键残基进行甲基标记,从而促进了对超分子颗粒的结构、动态和相互作用的研究。简要回顾了甲基标记方法,然后总结了三个不同分子机器的应用,以说明现在可以解决的问题类型。还强调了未来创新将带来进一步改进的领域。