膜蛋白结构研究方法新进展.pdfVIP

《分子生物物理学》研究生评述2003 膜蛋白结构研究方法新进展 姓名：金聪 学号 系别：免疫系 摘要 膜蛋白是一类结构独特的蛋白质，执行很多基本的和重要的细胞生物学功 能。了解膜蛋白在生物膜上的基本构象对研究膜蛋白的精细拓扑结构和功能具有 重要意义。但是膜蛋白的疏水特性使其需要与生物膜共同形成稳定的自然构象, 因此至今在蛋白质组学的研究中对膜蛋白所知甚少。了解分子结构是了解生物大 分子功能的一个重要途径，因此在这里对近来膜分子结构研究领域的进展做一个 简要的概述。晶体学方法，单颗粒方法和原子力显微镜为膜蛋白的研究提供了大 量的细节数据。固相核磁共振技术提供了决定跨膜 α螺旋结构的方向约束数据 和精确的分子间距离约束数据。直接位点标记的旋转电子顺磁共振可以得到更长 的距离约束数据，但是目前的标记策略仍然具有局限性。其他的方法也被用于分 析跨膜 α螺旋束，例如位点特异的红外二色性分析使得在脂双层中定向分析跨 膜 α螺旋，以及利用进化保守数据进行计算和预测的新工具的结合应用。 关键词：膜蛋白 三维结构 晶体学方法 核磁共振方法 结构预测 Abstract Membrane proteins carry out essential cellular functions. However， their hydrophobic nature makes them notoriously difficult to study，the task of determining the structure of membrane proteins has been hindered by experimental difficulties associated with their lipid-embedded domains，which keeping them consistently underrepresented in proteomic analyses. Here, we provide an overview of recently developed experimental and predictive tools that are changing our view of this largely unexplored territory of structural biology. Crystallography, single-particle methods and atomic force microscopy are being used to study huge membrane proteins with increasing detail. Solid-state nuclear magnetic resonance strategies provide orientational constraints for structure determination of transmembrane (TM) α-helices and accurate measurements of intramolecular distances, even in very complex systems. Longer distance constraints are determined by site-directed spin labelling electron paramagnetic resonance, but current labelling strategies still constitute some limitation. Other methods, such as site-specific infrared dichroism, enable orientational analysis of TM a-helices in aligned bilayers and, combined with novel computational and predictive tools that use evolutionary conservation data, are being used to analyze