Maxwell Karen L, Wildes David, Zarrine-Afsar Arash, De Los Rios Miguel A, Brown Andrew G, Friel Claire T, Hedberg Linda, Horng Jia-Cherng, Bona Diane, Miller Erik J, Vallée-Bélisle Alexis, Main Ewan R G, Bemporad Francesco, Qiu Linlin, Teilum Kaare, Vu Ngoc-Diep, Edwards Aled M, Ruczinski Ingo, Poulsen Flemming M, Kragelund Birthe B, Michnick Stephen W, Chiti Fabrizio, Bai Yawen, Hagen Stephen J, Serrano Luis, Oliveberg Mikael, Raleigh Daniel P, Wittung-Stafshede Pernilla, Radford Sheena E, Jackson Sophie E, Sosnick Tobin R, Marqusee Susan, Davidson Alan R, Plaxco Kevin W
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.
Protein Sci. 2005 Mar;14(3):602-16. doi: 10.1110/ps.041205405. Epub 2005 Feb 2.
Recent years have seen the publication of both empirical and theoretical relationships predicting the rates with which proteins fold. Our ability to test and refine these relationships has been limited, however, by a variety of difficulties associated with the comparison of folding and unfolding rates, thermodynamics, and structure across diverse sets of proteins. These difficulties include the wide, potentially confounding range of experimental conditions and methods employed to date and the difficulty of obtaining correct and complete sequence and structural details for the characterized constructs. The lack of a single approach to data analysis and error estimation, or even of a common set of units and reporting standards, further hinders comparative studies of folding. In an effort to overcome these problems, we define here a "consensus" set of experimental conditions (25 degrees C at pH 7.0, 50 mM buffer), data analysis methods, and data reporting standards that we hope will provide a benchmark for experimental studies. We take the first step in this initiative by describing the folding kinetics of 30 apparently two-state proteins or protein domains under the consensus conditions. The goal of our efforts is to set uniform standards for the experimental community and to initiate an accumulating, self-consistent data set that will aid ongoing efforts to understand the folding process.
近年来,已经发表了一些预测蛋白质折叠速率的经验关系和理论关系。然而,由于与不同蛋白质组的折叠和去折叠速率、热力学以及结构比较相关的各种困难,我们测试和完善这些关系的能力受到了限制。这些困难包括迄今为止所采用的实验条件和方法范围广泛且可能相互混淆,以及难以获得所表征构建体的正确和完整的序列及结构细节。缺乏单一的数据分析和误差估计方法,甚至缺乏一套通用的单位和报告标准,进一步阻碍了折叠的比较研究。为了克服这些问题,我们在此定义了一组“共识”实验条件(pH 7.0、50 mM缓冲液,25摄氏度)、数据分析方法和数据报告标准,我们希望这些能为实验研究提供一个基准。我们通过描述30种明显呈两态的蛋白质或蛋白质结构域在共识条件下的折叠动力学,迈出了这一倡议的第一步。我们努力的目标是为实验界设定统一标准,并启动一个不断积累、自洽的数据集,以帮助正在进行的理解折叠过程的努力。