Department of Anesthesiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
J Phys Chem B. 2010 Jun 10;114(22):7649-55. doi: 10.1021/jp1009675.
Neuronal nicotinic acetylcholine receptors (nAChRs) have been implicated as targets for general anesthetics, but the functional responses to anesthetic modulation vary considerably among different subtypes of nAChRs. Inhaled general anesthetics, such as halothane, could effectively inhibit the channel activity of the alpha4beta2 nAChR but not the homologous alpha7 nAChR. To understand why alpha7 is insensitive to inhaled general anesthetics, we performed multiple sets of 20 ns molecular dynamics (MD) simulations on the closed- and open-channel alpha7 in the absence and presence of halothane and critically compared the results with those from our studies on the alpha4beta2 nAChR (Liu et al. J. Phys. Chem. B 2009, 113, 12581 and Liu et al. J. Phys. Chem. B 2010, 114, 626). Several halothane binding sites with fairly high binding affinities were identified in both closed- and open-channel alpha7, suggesting that a lack of sensitive functional responses of the alpha7 nAChR to halothane in the previous experiments was unlikely due to a lack of halothane interaction with alpha7. The binding affinities of halothane in alpha7 seemed to be protein conformation-dependent. Overall, halothane affinity was higher in the closed-channel alpha7. Halothane binding to alpha7 did not induce profound changes in alpha7 structure and dynamics that could be related to the channel function. In contrast, correlated motion of the open-channel alpha4beta2 was reduced substantially in the presence of halothane, primarily due to the more susceptible nature of beta2 to anesthetic modulation. The amphiphilic extracellular and transmembrane domain interface of the beta2 subunit is attractive to halothane and is susceptible to halothane perturbation, which may be why alpha4beta2 is functionally more sensitive to halothane than alpha7.
神经元烟碱型乙酰胆碱受体 (nAChRs) 已被认为是全身麻醉剂的靶标,但不同亚型的 nAChR 对麻醉调制的功能反应差异很大。吸入性全身麻醉剂,如氟烷,可有效抑制α4β2 nAChR 的通道活性,但对同源α7 nAChR 则无效。为了了解为什么α7 对吸入性全身麻醉剂不敏感,我们在没有和存在氟烷的情况下对封闭和开放通道的α7 进行了多组 20ns 分子动力学 (MD) 模拟,并与我们在α4β2 nAChR 上的研究结果进行了严格比较(Liu 等人,J. Phys. Chem. B 2009, 113, 12581 和 Liu 等人,J. Phys. Chem. B 2010, 114, 626)。在封闭和开放通道的α7 中都鉴定出了几个具有相当高结合亲和力的氟烷结合位点,这表明以前的实验中α7 nAChR 对氟烷缺乏敏感的功能反应不太可能是由于缺乏氟烷与α7 的相互作用。α7 中氟烷的结合亲和力似乎取决于蛋白质构象。总体而言,封闭通道中的α7 中氟烷的亲和力更高。氟烷与α7 的结合不会引起α7 结构和动力学的深刻变化,这些变化可能与通道功能有关。相比之下,在氟烷存在下,开放通道的α4β2 的相关运动大大减少,主要是由于β2 对麻醉调制的敏感性更高。β2 亚基的亲脂性细胞外和跨膜域界面对氟烷有吸引力,并且容易受到氟烷的干扰,这可能是为什么α4β2 比α7 对氟烷的功能更敏感的原因。