Vermaas J V, Trebesch N, Mayne C G, Thangapandian S, Shekhar M, Mahinthichaichan P, Baylon J L, Jiang T, Wang Y, Muller M P, Shinn E, Zhao Z, Wen P-C, Tajkhorshid E
Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL, United States; University of Illinois at Urbana-Champaign, Urbana, IL, United States; Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, United States.
University of Illinois at Urbana-Champaign, Urbana, IL, United States; Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, United States.
Methods Enzymol. 2016;578:373-428. doi: 10.1016/bs.mie.2016.05.042. Epub 2016 Jul 11.
Membrane transporters mediate one of the most fundamental processes in biology. They are the main gatekeepers controlling active traffic of materials in a highly selective and regulated manner between different cellular compartments demarcated by biological membranes. At the heart of the mechanism of membrane transporters lie protein conformational changes of diverse forms and magnitudes, which closely mediate critical aspects of the transport process, most importantly the coordinated motions of remotely located gating elements and their tight coupling to chemical processes such as binding, unbinding and translocation of transported substrate and cotransported ions, ATP binding and hydrolysis, and other molecular events fueling uphill transport of the cargo. An increasing number of functional studies have established the active participation of lipids and other components of biological membranes in the function of transporters and other membrane proteins, often acting as major signaling and regulating elements. Understanding the mechanistic details of these molecular processes require methods that offer high spatial and temporal resolutions. Computational modeling and simulations technologies empowered by advanced sampling and free energy calculations have reached a sufficiently mature state to become an indispensable component of mechanistic studies of membrane transporters in their natural environment of the membrane. In this article, we provide an overview of a number of major computational protocols and techniques commonly used in membrane transporter modeling and simulation studies. The article also includes practical hints on effective use of these methods, critical perspectives on their strengths and weak points, and examples of their successful applications to membrane transporters, selected from the research performed in our own laboratory.
膜转运蛋白介导生物学中最基本的过程之一。它们是主要的守门人,以高度选择性和受调控的方式控制物质在由生物膜界定的不同细胞区室之间的主动运输。膜转运蛋白机制的核心是各种形式和幅度的蛋白质构象变化,这些变化紧密介导运输过程的关键方面,最重要的是远距离门控元件的协同运动及其与化学过程的紧密耦合,如被运输底物和共运输离子的结合、解离和转运、ATP结合和水解,以及其他推动货物上坡运输的分子事件。越来越多的功能研究证实,生物膜中的脂质和其他成分积极参与转运蛋白及其他膜蛋白的功能,它们常作为主要的信号和调节元件。理解这些分子过程的机制细节需要具备高空间和时间分辨率的方法。由先进的采样和自由能计算支持的计算建模和模拟技术已发展到足够成熟的阶段,成为在膜的自然环境中对膜转运蛋白进行机制研究不可或缺的组成部分。在本文中,我们概述了膜转运蛋白建模和模拟研究中常用的一些主要计算协议和技术。本文还包括有效使用这些方法的实用提示、对其优缺点的批判性观点,以及从我们自己实验室开展的研究中选取的它们成功应用于膜转运蛋白的实例。