Eftink M R
Department of Chemistry, University of Mississippi, University 38677.
Biophys J. 1994 Feb;66(2 Pt 1):482-501. doi: 10.1016/s0006-3495(94)80799-4.
This article discusses several strategies for the use steady-state and time-resolved fluorescence methods to monitor unfolding transitions in proteins. The assumptions and limitations of several methods are discussed. Simulations are presented to show that certain fluorescence observables directly track the population of states in an unfolding transition, whereas other observables skew the transition toward the dominant fluorescing species. Several examples are given, involving the unfolding of Staphylococcal aureus nuclease A, in which thermodynamic information is obtained for the temperature and denaturant induced transitions in this protein.
本文讨论了使用稳态和时间分辨荧光方法监测蛋白质展开转变的几种策略。文中讨论了几种方法的假设和局限性。通过模拟表明，某些荧光可观测量直接跟踪展开转变中状态的群体，而其他可观测量则使转变偏向占主导的荧光物种。文中给出了几个例子，涉及金黄色葡萄球菌核酸酶A的展开，从中获得了该蛋白质在温度和变性剂诱导转变中的热力学信息。
