Detergent-Free Membrane Protein Purification Using SMA Polymer.
Detergent-Free Membrane Protein Purification Using SMA Polymer.
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DOI：
10.1007/978-1-0716-2368-8_21
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发表时间：
2022-01-01
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影响因子：
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通讯作者：
Rothnie, Alice J
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文献类型：
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作者：
Broadbent, Luke;Depping, Peer;Rothnie, Alice J
研究膜蛋白的结构和功能的最大挑战之一是需要从膜中提取它们。传统上，这是使用去污剂来实现的，去污剂破坏膜并在蛋白质周围形成胶束，但这可能导致蛋白质功能和/或稳定性的问题。2009年，报道了一种替代方法，使用苯乙烯马来酸（SMA）共聚物提取由聚合物包封的脂质双层的小圆盘，称为SMALP（SMA脂质颗粒）。从那时起，这种方法已被证明适用于来自许多不同表达系统的一系列不同蛋白质。它允许提取和纯化靶蛋白，同时保持脂质双层环境。最近，这导致了几个新的高分辨率结构和新的见解功能。与任何方法一样，有一些限制和问题需要注意。在这里，我们描述了一个标准的协议，用于制备的聚合物和其用于膜蛋白纯化，还包括可能遇到的典型挑战的细节和可能的方法来解决这些问题。
One of the big challenges for the study of structure and function of membrane proteins is the need to extract them from the membrane. Traditionally this was achieved using detergents which disrupt the membrane and form a micelle around the protein, but this can cause issues with protein function and/or stability. In 2009 an alternative approach was reported, using styrene maleic acid (SMA) copolymer to extract small discs of lipid bilayer encapsulated by the polymer and termed SMALPs (SMA lipid particles). Since then this approach has been shown to work for a range of different proteins from many different expression systems. It allows the extraction and purification of a target protein while maintaining a lipid bilayer environment. Recently this has led to several new high-resolution structures and novel insights to function. As with any method there are some limitations and issues to be aware of. Here we describe a standard protocol for preparation of the polymer and its use for membrane protein purification, and also include details of typical challenges that may be encountered and possible ways to address those.