Betaneli Viktoria, Schwille Petra
Biophysics, BIOTEC, Technische Universitat Dresden, Dresden, Germany.
Methods Mol Biol. 2013;974:253-78. doi: 10.1007/978-1-62703-275-9_12.
Fluorescence correlation spectroscopy (FCS) is a versatile technique to study membrane dynamics and protein-lipid interactions. It can provide information about diffusion coefficients, concentrations, and molecular interactions of proteins and lipids in the membrane. These parameters allow the determination of protein partitioning into different lipid environments, the identification of lipid domains, and the detection of lipid-protein complexes on the membrane. During the last decade, FCS studies were successfully performed on model membrane systems as also on living cells, to characterize protein-lipid interactions. Recent developments of the method described here improved quantitative measurements on membranes and decreased the number of potential artifacts. The aim of this chapter is to provide the reader with the necessary information and some practical guidelines to perform FCS studies on artificial and cellular membranes.
荧光相关光谱法(FCS)是一种用于研究膜动力学和蛋白质-脂质相互作用的通用技术。它可以提供有关膜中蛋白质和脂质的扩散系数、浓度以及分子相互作用的信息。这些参数有助于确定蛋白质在不同脂质环境中的分配情况、脂质结构域的识别以及膜上脂质-蛋白质复合物的检测。在过去十年中,FCS研究已成功应用于模型膜系统以及活细胞,以表征蛋白质-脂质相互作用。此处所述方法的最新进展改进了对膜的定量测量,并减少了潜在假象的数量。本章的目的是为读者提供进行人工膜和细胞膜FCS研究所需的信息及一些实用指南。