Robertson Janice L, Palmer Lawrence G, Roux Benoît
Program in Physiology, Biophysics and Systems Biology, Weill Graduate School of Medical Sciences, Cornell University, New York, NY 10065, USA.
J Gen Physiol. 2008 Dec;132(6):613-32. doi: 10.1085/jgp.200810068. Epub 2008 Nov 10.
Inward-rectifier potassium (Kir) channels differ from the canonical K(+) channel structure in that they possess a long extended pore (approximately 85 A) for ion conduction that reaches deeply into the cytoplasm. This unique structural feature is presumably involved in regulating functional properties specific to Kir channels, such as conductance, rectification block, and ligand-dependent gating. To elucidate the underpinnings of these functional roles, we examine the electrostatics of an ion along this extended pore. Homology models are constructed based on the open-state model of KirBac1.1 for four mammalian Kir channels: Kir1.1/ROMK, Kir2.1/IRK, Kir3.1/GIRK, and Kir6.2/KATP. By solving the Poisson-Boltzmann equation, the electrostatic free energy of a K(+) ion is determined along each pore, revealing that mammalian Kir channels provide a favorable environment for cations and suggesting the existence of high-density regions in the cytoplasmic domain and cavity. The contribution from the reaction field (the self-energy arising from the dielectric polarization induced by the ion's charge in the complex geometry of the pore) is unfavorable inside the long pore. However, this is well compensated by the electrostatic interaction with the static field arising from the protein charges and shielded by the dielectric surrounding. Decomposition of the static field provides a list of residues that display remarkable correspondence with existing mutagenesis data identifying amino acids that affect conduction and rectification. Many of these residues demonstrate interactions with the ion over long distances, up to 40 A, suggesting that mutations potentially affect ion or blocker energetics over the entire pore. These results provide a foundation for understanding ion interactions in Kir channels and extend to the study of ion permeation, block, and gating in long, cation-specific pores.
内向整流钾(Kir)通道与典型的钾通道结构不同,因为它们具有一个长的延伸孔道(约85埃)用于离子传导,该孔道深入细胞质。这种独特的结构特征可能参与调节Kir通道特有的功能特性,如电导、整流阻滞和配体依赖性门控。为了阐明这些功能作用的基础,我们研究了沿着这个延伸孔道的离子静电学。基于KirBac1.1的开放态模型构建了四种哺乳动物Kir通道的同源模型:Kir1.1/ROMK、Kir2.1/IRK、Kir3.1/GIRK和Kir6.2/KATP。通过求解泊松-玻尔兹曼方程,确定了钾离子沿着每个孔道的静电自由能,这表明哺乳动物Kir通道为阳离子提供了一个有利的环境,并暗示在细胞质区域和腔内存在高密度区域。反应场的贡献(由离子电荷在孔道复杂几何结构中诱导的介电极化产生的自能)在长孔道内部是不利的。然而,这被与蛋白质电荷产生的静电场的相互作用很好地补偿,并被周围的电介质屏蔽。静电场的分解提供了一系列残基,这些残基与现有的诱变数据显示出显著的对应关系,这些数据确定了影响传导和整流的氨基酸。这些残基中的许多在长达40埃的距离上与离子相互作用,这表明突变可能会影响整个孔道上的离子或阻滞剂能量学。这些结果为理解Kir通道中的离子相互作用提供了基础,并扩展到对长的、阳离子特异性孔道中的离子渗透、阻滞和门控的研究。