Findlay Heather E, Booth Paula J
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol, UK.
Methods Mol Biol. 2013;1063:117-24. doi: 10.1007/978-1-62703-583-5_6.
The native environment of integral membrane proteins is a highly complex lipid bilayer composed of many different types of lipids, the physical characteristics of which can profoundly influence protein stability, folding, and function. Secondary transporters are a class of protein where changes to both structure and activity have been observed in different bilayer environments. In order to study these interactions in vitro, it is necessary to extract and purify the protein and exchange it into an artificial lipid system that can be manipulated to control protein behavior. Liposomes are a commonly used model system that is particularly suitable for studying transporters. GalP and LacY can be reconstituted or refolded into vesicles with a high degree of efficiency for further structural analysis. Circular dichroism spectroscopy is an important technique in monitoring protein folding, which allows the decomposition of spectra into secondary structural components.
整合膜蛋白的天然环境是一个高度复杂的脂质双层，由许多不同类型的脂质组成，其物理特性可深刻影响蛋白质的稳定性、折叠和功能。次级转运蛋白是一类在不同双层环境中结构和活性均发生变化的蛋白质。为了在体外研究这些相互作用，有必要提取和纯化蛋白质，并将其交换到可操控以控制蛋白质行为的人工脂质体系中。脂质体是一种常用的模型体系，特别适合用于研究转运蛋白。GalP和LacY可以高效地重构成或重新折叠到囊泡中，以进行进一步的结构分析。圆二色光谱法是监测蛋白质折叠的一项重要技术，它能将光谱分解为二级结构成分。
