Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio, United States of America.
Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, Ohio, United States of America.
PLoS Comput Biol. 2022 Sep 9;18(9):e1010036. doi: 10.1371/journal.pcbi.1010036. eCollection 2022 Sep.
Intrinsically disordered proteins (IDPs) are highly dynamic systems that play an important role in cell signaling processes and their misfunction often causes human disease. Proper understanding of IDP function not only requires the realistic characterization of their three-dimensional conformational ensembles at atomic-level resolution but also of the time scales of interconversion between their conformational substates. Large sets of experimental data are often used in combination with molecular modeling to restrain or bias models to improve agreement with experiment. It is shown here for the N-terminal transactivation domain of p53 (p53TAD) and Pup, which are two IDPs that fold upon binding to their targets, how the latest advancements in molecular dynamics (MD) simulations methodology produces native conformational ensembles by combining replica exchange with series of microsecond MD simulations. They closely reproduce experimental data at the global conformational ensemble level, in terms of the distribution properties of the radius of gyration tensor, and at the local level, in terms of NMR properties including 15N spin relaxation, without the need for reweighting. Further inspection revealed that 10-20% of the individual MD trajectories display the formation of secondary structures not observed in the experimental NMR data. The IDP ensembles were analyzed by graph theory to identify dominant inter-residue contact clusters and characteristic amino-acid contact propensities. These findings indicate that modern MD force fields with residue-specific backbone potentials can produce highly realistic IDP ensembles sampling a hierarchy of nano- and picosecond time scales providing new insights into their biological function.
无规卷曲蛋白质(IDPs)是高度动态的系统,在细胞信号转导过程中发挥着重要作用,其功能障碍常常导致人类疾病。正确理解 IDP 的功能不仅需要在原子分辨率水平上对其三维构象集合进行真实的描述,还需要对其构象亚态之间的转换时间尺度进行描述。通常结合使用大量的实验数据和分子建模,通过限制或偏向模型来提高与实验的一致性。本文以 p53(p53TAD)和 Pup 的 N 端转录激活结构域为例,这两个 IDP 在与靶标结合时会发生折叠,展示了最新的分子动力学(MD)模拟方法如何通过结合 replica exchange 与一系列微秒 MD 模拟来产生天然构象集合。它们在全局构象集合水平上非常接近实验数据,在回转半径张量分布特性方面,以及在局部水平上,在 NMR 特性方面,包括 15N 自旋弛豫,而无需重新加权。进一步的检查表明,10-20%的单个 MD 轨迹显示出形成了实验 NMR 数据中未观察到的二级结构。通过图论对 IDP 集合进行了分析,以确定主要的残基间接触簇和特征氨基酸接触倾向。这些发现表明,具有残基特异性骨架势的现代 MD 力场可以产生高度真实的 IDP 集合,这些集合可以在纳米和皮秒时间尺度上进行采样,从而为它们的生物学功能提供新的见解。