Balbach J, Forge V, Lau W S, van Nuland N A, Brew K, Dobson C M
Oxford Centre for Molecular Sciences, New Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QT, UK.
Science. 1996 Nov 15;274(5290):1161-3. doi: 10.1126/science.274.5290.1161.
An approach is described to monitor directly at the level of individual residues the formation of structure during protein folding. A two-dimensional heteronuclear nuclear magnetic resonance (NMR) spectrum was recorded after the rapid initiation of the refolding of a protein labeled with nitrogen-15. The intensities and line shapes of the cross peaks in the spectrum reflected the kinetic time course of the folding events that occurred during the spectral accumulation. The method was used to demonstrate the cooperative nature of the acquisition of the native main chain fold of apo bovine alpha-lactalbumin. The general approach, however, should be applicable to the investigation of a wide range of chemical reactions.
本文描述了一种在蛋白质折叠过程中直接在单个残基水平监测结构形成的方法。在用氮-15标记的蛋白质快速重折叠开始后，记录了二维异核核磁共振（NMR）谱。谱中交叉峰的强度和线形反映了谱累积期间发生的折叠事件的动力学时间进程。该方法用于证明脱辅基牛α-乳白蛋白天然主链折叠形成的协同性质。然而，这种通用方法应适用于广泛化学反应的研究。
