Coronel Lucia, Muccio Giovanni Di, Rothberg Brad, Giacomello Alberto, Carnevale Vincenzo
ArXiv. 2024 May 7:arXiv:2405.04644v1.
The large-conductance, calcium-activated potassium (BK) channel lacks the typical intracellular bundle-crossing gate present in most ion channels of the 6TM family. This observation, initially inferred from Ca$^{2+}$-free-pore accessibility experiments and recently corroborated by a CryoEM structure of the non-conductive state, raises a puzzling question: how can gating occur in absence of steric hindrance? To answer this question, we carried out molecular simulations and accurate free energy calculations to obtain a microscopic picture of the sequence of events that, starting from a Ca$^{2+}$-free state leads to ion conduction upon Ca$^{2+}$ binding. Our results highlight an unexpected role for annular lipids, which turn out to be an integral part of the gating machinery. Due to the presence of fenestrations, the "closed" Ca$^{2+}$-free pore can be occupied by the methyl groups from the lipid alkyl chains. This dynamic occupancy triggers and stabilizes the nucleation of a vapor bubble into the inner pore cavity, thus hindering ion conduction. By contrast, Ca$^{2+}$ binding results into a displacement of these lipids outside the inner cavity, lowering the hydrophobicity of this region and thus allowing for pore hydration and conduction. This lipid-mediated hydrophobic gating rationalizes several seemingly problematic experimental observations, including the state-dependent pore accessibility of blockers.
大电导钙激活钾(BK)通道缺乏大多数6TM家族离子通道中存在的典型细胞内束交叉门控。这一观察结果最初是从无Ca²⁺时孔道可及性实验推断出来的,最近通过非传导状态的冷冻电镜结构得到了证实,这就提出了一个令人困惑的问题:在没有空间位阻的情况下,门控是如何发生的?为了回答这个问题,我们进行了分子模拟和精确的自由能计算,以获得从无Ca²⁺状态开始,在Ca²⁺结合后导致离子传导的一系列事件的微观图景。我们的结果突出了环状脂质的一个意想不到的作用,结果表明它们是门控机制的一个组成部分。由于存在小孔,“关闭”的无Ca²⁺孔道可以被脂质烷基链上的甲基占据。这种动态占据触发并稳定了内孔腔中蒸汽泡的成核,从而阻碍了离子传导。相比之下,Ca²⁺结合导致这些脂质向内腔外位移,降低了该区域的疏水性,从而允许孔道水合和传导。这种脂质介导的疏水门控解释了几个看似有问题的实验观察结果,包括阻滞剂的状态依赖性孔道可及性。