Goddard Alan D, Dijkman Patricia M, Adamson Roslin J, dos Reis Rosana Inácio, Watts Anthony
School of Life Sciences, University of Lincoln, Lincoln, United Kingdom.
Biomembrane Structure Unit, Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
Methods Enzymol. 2015;556:405-24. doi: 10.1016/bs.mie.2015.01.004. Epub 2015 Mar 20.
Membrane proteins are the gatekeepers to the cell and are essential to the function of all cells, controlling the flow of molecules and information across the cell membrane. Much effort has been put into the development of systems for studying membrane proteins in simplified environments that nevertheless mimic their native lipid environment. After isolation and production of purified membrane proteins in detergent, it is often necessary to reconstitute them into a lipid structure such as liposome, nanodisc, or lipodisq. Each of these has the advantage of returning the protein to a defined lipid environment, and the choice of system depends on the application. Regardless of the system to be used, the fundamental process involves the removal of detergent and incorporation of the protein into a stable lipid system. This chapter details methodologies we have developed, mainly focussed on the model G protein-coupled receptor (GPCR) neurotensin receptor 1, and the GPCR-homologue and model, bacteriorhopdopsin.
膜蛋白是细胞的守门人，对所有细胞的功能至关重要，控制着分子和信息跨细胞膜的流动。人们付出了很多努力来开发在简化环境中研究膜蛋白的系统，这些环境仍能模拟其天然脂质环境。在去污剂中分离并生产纯化的膜蛋白后，通常需要将它们重构到脂质结构中，如脂质体、纳米盘或脂质盘。这些方法中的每一种都有将蛋白质恢复到确定脂质环境的优点，系统的选择取决于应用。无论使用何种系统，基本过程都包括去除去污剂并将蛋白质掺入稳定的脂质系统中。本章详细介绍了我们开发的方法，主要集中在模型G蛋白偶联受体（GPCR）神经降压素受体1以及GPCR同源物和模型细菌视紫红质上。
