Seddon Annela M, Curnow Paul, Booth Paula J
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK.
Biochim Biophys Acta. 2004 Nov 3;1666(1-2):105-17. doi: 10.1016/j.bbamem.2004.04.011.
Studying membrane proteins represents a major challenge in protein biochemistry, with one of the major difficulties being the problems encountered when working outside the natural lipid environment. In vitro studies such as crystallization are reliant on the successful solubilization or reconstitution of membrane proteins, which generally involves the careful selection of solubilizing detergents and mixed lipid/detergent systems. This review will concentrate on the methods currently available for efficient reconstitution and solubilization of membrane proteins through the use of detergent micelles, mixed lipid/detergent micelles and bicelles or liposomes. We focus on the relevant molecular properties of the detergents and lipids that aid understanding of these processes. A significant barrier to membrane protein research is retaining the stability and function of the protein during solubilization, reconstitution and crystallization. We highlight some of the lessons learnt from studies of membrane protein folding in vitro and give an overview of the role that lipids can play in stabilizing the proteins.
研究膜蛋白是蛋白质生物化学中的一项重大挑战,其中一个主要困难是在天然脂质环境之外进行研究时遇到的问题。诸如结晶等体外研究依赖于膜蛋白的成功溶解或重构,这通常涉及仔细选择增溶去污剂以及混合脂质/去污剂体系。本综述将集中讨论目前通过使用去污剂胶束、混合脂质/去污剂胶束、双分子层或脂质体来有效重构和溶解膜蛋白的方法。我们关注有助于理解这些过程的去污剂和脂质的相关分子特性。膜蛋白研究的一个重大障碍是在溶解、重构和结晶过程中保持蛋白质的稳定性和功能。我们强调了从体外膜蛋白折叠研究中吸取的一些经验教训,并概述了脂质在稳定蛋白质方面可以发挥的作用。