A Transient and Low-Populated Protein-Folding Intermediate at Atomic Resolution
A Transient and Low-Populated Protein-Folding Intermediate at Atomic Resolution
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DOI：
10.1126/science.1191723
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发表时间：
2010-09-10
期刊：
影响因子：
56.9
通讯作者：
Kay, Lewis E.
中科院分区：
文献类型：
--
作者：
Korzhnev, Dmitry M.;Religa, Tomasz L.;Kay, Lewis E.
蛋白质可以采样对功能至关重要的构象状态，但由于它们的低占有率和短寿命而很少被直接检测到。在这项工作中，我们使用的化学位移和键矢量取向的限制，从核磁共振弛豫色散光谱，与化学位移为基础的方法进行结构说明，以确定一个原子分辨率的结构的“看不见的”折叠中间的一个小蛋白质模块：FF域。该结构揭示了非天然元素，防止在蛋白质的羧基末端部分形成天然构象。这与折叠的动力学一致，其中结构良好的中间体快速形成，然后缓慢重排至天然状态。该方法介绍了一个一般的策略，低人口和瞬时形成的蛋白质状态的结构测定。
Proteins can sample conformational states that are critical for function but are seldom detected directly because of their low occupancies and short lifetimes. In this work, we used chemical shifts and bond-vector orientation constraints obtained from nuclear magnetic resonance relaxation dispersion spectroscopy, in concert with a chemical shift-based method for structure elucidation, to determine an atomic-resolution structure of an "invisible" folding intermediate of a small protein module: the FF domain. The structure reveals non-native elements preventing formation of the native conformation in the carboxyl-terminal part of the protein. This is consistent with the kinetics of folding in which a well-structured intermediate forms rapidly and then rearranges slowly to the native state. The approach introduces a general strategy for structure determination of low-populated and transiently formed protein states.