CERM, University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy.
J Am Chem Soc. 2010 Sep 29;132(38):13553-8. doi: 10.1021/ja1063923.
The concept of maximum occurrence (MO), i.e., the maximum percent of time that flexible proteins can spend in any given conformation, is introduced, and a rigorous method is developed to extensively sample the conformational space and to construct MO maps from experimental data. The method is tested in a case study, the flexible two-domain protein calmodulin (CaM), using SAXS and NMR data (i.e., pseudocontact shifts and self-orientation residual dipolar couplings arising from the presence of paramagnetic lanthanide ions), revealing that the "closed" and "fully extended" conformations trapped in the crystalline forms of CaM have MOs of only 5 and 15%, respectively. Compact conformations in general have small MOs, whereas some extended conformations have MO as high as 35%, strongly suggesting these conformations to be most abundant in solution. The method is universally applicable as it requires only standard SAXS data and specific NMR data on lanthanide derivatives of the protein (using native metal sites or lanthanide tagging). The computer program is publicly available using the grid computing infrastructure through the authors' Web portal.
引入最大出现概率（MO）的概念，即柔性蛋白质在任何给定构象中可以花费的最大时间百分比。开发了一种严格的方法来广泛采样构象空间，并从实验数据构建 MO 图谱。该方法在一个案例研究中得到了测试，该案例研究是柔性双域蛋白钙调蛋白（CaM），使用 SAXS 和 NMR 数据（即来自顺磁镧系离子存在的赝接触位移和自取向剩余偶极耦合），结果表明，在 CaM 的晶体形式中捕获的“封闭”和“完全伸展”构象的 MO 分别只有 5%和 15%。一般来说，紧凑构象的 MO 较小，而一些伸展构象的 MO 高达 35%，这强烈表明这些构象在溶液中最丰富。该方法具有普遍性，因为它只需要标准的 SAXS 数据和蛋白质的镧系衍生物的特定 NMR 数据（使用天然金属位点或镧系元素标记）。该计算机程序可通过作者的 Web 门户使用网格计算基础设施公开使用。
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