Tikhonov D B, Zhorov B S
Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg.
Biophys J. 1998 Jan;74(1):242-55. doi: 10.1016/S0006-3495(98)77783-5.
A model of the nicotinic acetylcholine receptor ion channel was elaborated based on the data from electron microscopy, affinity labeling, cysteine scanning, mutagenesis studies, and channel blockade. A restrained Monte Carlo minimization method was used for the calculations. Five identical M2 segments (the sequence EKMTLSISVL10LALTVFLLVI20V) were arranged in five-helix bundles with various geometrical profiles of the pore. For each bundle, energy profiles for chlorpromazine, QX-222, pentamethonium, and other blocking drugs pulled through the pore were calculated. An optimal model obtained allows all of the blockers free access to the pore, but retards them at the rings of residues known to contribute to the corresponding binding sites. In this model, M2 helices are necessarily kinked. They come into contact with each other at the cytoplasmic end but diverge at the synaptic end, where N-termini of M1 segments may contribute to the pore. The kinks disengage alpha-helical H-bonds between Ala12 and Ser8. The uncoupled lone electron pairs of Ser8 carbonyl oxygens protrude into the pore, forming a hydrophilic ring that may be important for the permeation of cations. A split network of H-bonds provides a flexibility to the chains Val9-Ala12, the numerous conformations of which form only two or three intrasegment H-bonds. The cross-ectional dimensions of the interface between the flexible chains vary essentially at the level of Leu11. We suggest that conformational transitions in the chains Val9-Ala12 are responsible for the channel gating, whereas rotations of more stable alpha-helical parts of M2 segments may be necessary to transfer the channel in the desensitized state.
基于电子显微镜、亲和标记、半胱氨酸扫描、诱变研究和通道阻断的数据,构建了烟碱型乙酰胆碱受体离子通道模型。计算采用了受限蒙特卡罗最小化方法。五个相同的M2片段(序列为EKMTLSISVL10LALTVFLLVI20V)排列成具有不同孔几何形状的五螺旋束。对于每个束,计算了氯丙嗪、QX - 222、五甲铵和其他阻断药物通过孔时的能量分布。获得的最佳模型允许所有阻断剂自由进入孔,但在已知有助于相应结合位点的残基环处减缓它们的进入。在该模型中,M2螺旋必然是扭结的。它们在细胞质端相互接触,但在突触端发散,M1片段的N端可能对孔有贡献。扭结使Ala12和Ser8之间的α - 螺旋氢键断开。Ser8羰基氧的未偶联孤对电子伸入孔中,形成一个亲水环,这可能对阳离子的通透很重要。氢键的分裂网络为Val9 - Ala12链提供了灵活性,其众多构象仅形成两三个链内氢键。柔性链之间界面的横截面尺寸在Leu11水平上有很大变化。我们认为Val9 - Ala12链中的构象转变负责通道门控,而M2片段更稳定的α - 螺旋部分的旋转可能是将通道转变为脱敏状态所必需的。