Vallurupalli Pramodh, Hansen D Flemming, Kay Lewis E
Departments of Molecular Genetics, Biochemistry, and Chemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
J Am Chem Soc. 2008 Mar 5;130(9):2734-5. doi: 10.1021/ja710817g. Epub 2008 Feb 8.
A general method for obtaining quantitative structural information on invisible, excited protein states by solution-based NMR spectroscopy is presented. The approach exploits relaxation dispersion techniques in which changes in chemical shifts between ground and excited states are monitored in solutions with and without small amounts of residual molecular alignment. This allows the calculation of differences in chemical shifts induced by alignment that can be directly related to molecular structure, in cases where the orientation and magnitude of the chemical-shift tensor are well defined. An example using carbonyl chemical shifts as probes of a protein-ligand binding reaction is presented to illustrate and validate the method.
本文介绍了一种通过基于溶液的核磁共振光谱法获取关于不可见激发态蛋白质定量结构信息的通用方法。该方法利用弛豫色散技术，在有无少量残余分子排列的溶液中监测基态和激发态之间化学位移的变化。在化学位移张量的方向和大小定义明确的情况下，这允许计算由排列引起的化学位移差异，而这些差异可直接与分子结构相关。文中给出了一个使用羰基化学位移作为蛋白质 - 配体结合反应探针的例子，以说明和验证该方法。
