Research School of Biology, Australian National University, Canberra, Australia.
Biophys J. 2011 Dec 7;101(11):2671-8. doi: 10.1016/j.bpj.2011.10.045.
Using both Brownian and molecular dynamics, we replicate many of the salient features of Kv1.2, including the current-voltage-concentration profiles and the binding affinity and binding mechanisms of charybdotoxin, a scorpion venom. We also elucidate how structural differences in the inner vestibule can give rise to significant differences in its permeation characteristics. Current-voltage-concentration profiles are constructed using Brownian dynamics simulations, based on the crystal structure 2A79. The results are compatible with experimental data, showing similar conductance, rectification, and saturation with current. Unlike KcsA, for example, the inner pore of Kv1.2 is mainly hydrophobic and neutral, and to explore the consequences of this, we investigate the effect of mutating neutral proline residues at the mouth of the inner vestibule to charged aspartate residues. We find an increased conductance, less inward rectification, and quicker saturation of the current-voltage profile. Our simulations use modifications to our Brownian dynamics program that extend the range of channels that can be usefully modeled. Using molecular dynamics, we investigate the binding of the charybdotoxin scorpion venom to the outer vestibule of the channel. A potential of mean force is derived using umbrella sampling, giving a dissociation constant within a factor of ∼2 to experimentally derived constants. The residues involved in the toxin binding are in agreement with experimental mutagenesis studies. We thus show that the experimental observations on the voltage-gated channel, including the toxin-channel interaction, can reliably be replicated by using the two widely used computational tools.
我们使用布朗动力学和分子动力学,复制了 Kv1.2 的许多显著特征,包括电流-电压-浓度曲线以及 scorpion venom 毒素 charybdotoxin 的结合亲和力和结合机制。我们还阐明了内部前庭结构的差异如何导致其渗透特性的显著差异。电流-电压-浓度曲线是基于晶体结构 2A79 通过布朗动力学模拟构建的。结果与实验数据相符,显示出相似的电导、整流和电流饱和。与 KcsA 不同,例如,Kv1.2 的内部孔隙主要是疏水和中性的,为了探究这一结果,我们研究了将内部前庭入口处的中性脯氨酸残基突变为带电荷的天冬氨酸残基的影响。我们发现电导增加,内向整流减少,电流-电压曲线的饱和更快。我们的模拟使用了对布朗动力学程序的修改,扩展了可以有效建模的通道范围。我们使用分子动力学研究了 charybdotoxin 毒素与通道外前庭的结合。使用伞状采样得出平均力势,得到的离解常数与实验得出的常数相差约 2 倍。与毒素结合的残基与实验诱变研究一致。因此,我们表明,包括毒素-通道相互作用在内的电压门控通道的实验观察可以通过使用两种广泛使用的计算工具可靠地复制。