Odera Mitsuhiko, Furuta Tadaomi, Sohma Yoshiro, Sakurai Minoru
Center for Biological Resources and Informatics, Tokyo Institute of Technology, Yokohama, Kanagawa 226-8501, Japan.
Department of Pharmaceutical Sciences, Graduate School of Pharmacy and Center for Medical Science, International University of Health and Welfare, Ohtawara, Tochigi 324-8501, Japan.
Biophys Physicobiol. 2018 Feb 6;15:33-44. doi: 10.2142/biophysico.15.0_33. eCollection 2018.
Cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel that belongs to the ATP binding cassette protein superfamily. Deletion of phenylalanine at position 508 (ΔF508) is the most common CF-associated mutation and is present in nearly 90% of CF patients. Currently, atomistic level studies are insufficient for understanding the mechanism by which the deletion of a single amino acid causes greatly reduced folding as well as trafficking and gating defects. To clarify this mechanism, we first constructed an atomic model of the inward-facing ΔF508-CFTR and performed allatom molecular dynamics (MD) simulations of the protein in a membrane environment. All of the computational methodologies used are based on those developed in our previous study for wild-type CFTR. Two important findings were obtained. First, consistent with several previous computational results, the deletion of F508 causes a disruption of a hydrophobic cluster located at the interface between the nucleotide binding domain 1 (NBD1) and intracellular loop 4 (ICL4). This exerts unfavorable influences on the correlated motion between ICLs and transmembrane domains (TMDs), likely resulting in gating defects. Second, the F508 deletion affected the NBD1-NBD2 interface via allosteric communication originating from the correlated motion between NBDs and ICLs. As a result, several unusual inter-residue interactions are caused at the NBD1-NBD2 interface. In other words, their correct dimerization is impaired. This study provided insight into the atomic-level details of structural and dynamics changes caused by the ΔF508 mutation and thus provides good insight for drug design.
囊性纤维化跨膜传导调节因子(CFTR)是一种阴离子通道,属于ATP结合盒蛋白超家族。第508位苯丙氨酸缺失(ΔF508)是最常见的与囊性纤维化相关的突变,近90%的囊性纤维化患者存在该突变。目前,原子水平的研究不足以理解单个氨基酸缺失导致折叠大幅减少以及转运和门控缺陷的机制。为了阐明这一机制,我们首先构建了向内的ΔF508 - CFTR原子模型,并在膜环境中对该蛋白进行了全原子分子动力学(MD)模拟。所有使用的计算方法均基于我们之前针对野生型CFTR开发的方法。获得了两个重要发现。首先,与之前的几个计算结果一致,F508的缺失导致位于核苷酸结合结构域1(NBD1)和细胞内环4(ICL4)之间界面的疏水簇破坏。这对ICL和跨膜结构域(TMD)之间的相关运动产生不利影响,可能导致门控缺陷。其次,F508缺失通过源自NBD和ICL之间相关运动的变构通讯影响NBD1 - NBD2界面。结果,在NBD1 - NBD2界面引起了几种异常的残基间相互作用。换句话说,它们的正确二聚化受到损害。本研究深入了解了由ΔF508突变引起的结构和动力学变化的原子水平细节,从而为药物设计提供了很好的见解。