University of Toronto, Department of Molecular Genetics, 1 King's College Circle, Toronto, Ontario, Canada.
J Am Chem Soc. 2010 Jun 9;132(22):7589-91. doi: 10.1021/ja102090z.
A simple method is presented for quantifying Ile chi(2) rotamer distributions in proteins based on the measurement of Ile (13)C(delta1) chemical shifts. The methodology is well suited for applications involving very high molecular weight protein complexes, where other NMR parameters such as side-chain scalar coupling constants that report on dihedral angles cannot be measured or for studies of invisible, excited protein states, where chemical shifts are obtained from analysis of CPMG relaxation dispersion profiles. The utility of the approach is demonstrated by an application to the folding reaction of a mutant Fyn SH3 domain, where Ile side-chain structure and dynamics of an on-folding pathway intermediate state are studied.
本文提出了一种基于测量异亮氨酸 (13)C(delta1) 化学位移来量化蛋白质中异亮氨酸 chi(2) 构象分布的简单方法。该方法非常适合应用于涉及非常高分子量蛋白质复合物的情况，在这种情况下，无法测量报告二面角的其他 NMR 参数，如侧链标量耦合常数，或者用于研究不可见的、激发态蛋白质，在这种情况下，通过分析 CPMG 弛豫弥散曲线可以获得化学位移。该方法的实用性通过对突变 Fyn SH3 结构域折叠反应的应用得到了证明，其中研究了折叠途径中间态异亮氨酸侧链结构和动力学。
