Huang Xiaoqin, Dong Feng, Zhou Huan-Xiang
Institute of Molecular Biophysics and School of Computational Science, Department of Physics, Florida State University, Tallahassee, FL 32306, USA.
J Am Chem Soc. 2005 May 11;127(18):6836-49. doi: 10.1021/ja042641q.
Brownian dynamics (BD) and molecular dynamics (MD) simulations and electrostatic calculations were performed to study the binding process of kappa-PVIIA to the Shaker potassium channel and the structure of the resulting complex. BD simulations, guided by electrostatic interactions, led to an initial alignment between the toxin and the channel protein. MD simulations were then carried out to allow for rearrangements from this initial structure. After approximately 4 ns, a critical "induced fit" process was observed to last for approximately 2 ns. In this process, the interface was reorganized, and side chains were moved so that favorable atomic contacts were formed or strengthened, while unfavorable contacts were eliminated. The final complex structure was stabilized through electrostatic interactions with the positively charged side chain of Lys7 of kappa-PVIIA deeply inserted into the channel pore and other hydrogen bonds and by hydrophobic interactions involving Phe9 and Phe23 of the toxin. The validity of the predicted structure for the complex was assessed by calculating the effects of mutating charged and polar residues of both the toxin and the channel protein, with the calculated effects correlating reasonably well with experimental data. The present study suggests a general binding mechanism, whereby proteins are pre-aligned in their diffusional encounter by long-range electrostatic attraction, and nanosecond-scale rearrangements within the initial complex then lead to a specifically bound complex.
进行了布朗动力学(BD)和分子动力学(MD)模拟以及静电计算,以研究κ-PVIIA与Shaker钾通道的结合过程以及所得复合物的结构。在静电相互作用的引导下,BD模拟使毒素与通道蛋白之间实现了初始排列。随后进行MD模拟,以允许从该初始结构进行重排。大约4纳秒后,观察到一个关键的“诱导契合”过程持续了约2纳秒。在此过程中,界面发生重组,侧链移动,从而形成或加强了有利的原子接触,同时消除了不利的接触。最终的复合物结构通过与深深插入通道孔中的κ-PVIIA的Lys7带正电荷侧链的静电相互作用以及其他氢键以及涉及毒素的Phe9和Phe23的疏水相互作用而得以稳定。通过计算毒素和通道蛋白的带电和极性残基突变的影响,评估了复合物预测结构的有效性,计算结果与实验数据具有合理的相关性。本研究提出了一种普遍的结合机制,即蛋白质在扩散相遇时通过长程静电吸引进行预排列,然后初始复合物内的纳秒级重排导致形成特异性结合的复合物。