Hink Mark A, Postma Marten
Section of Molecular Cytology, van Leeuwenhoek Centre for Advanced Microscopy (LCAM), Swammerdam Institute for Life Sciences (SILS), University of Amsterdam, The Netherlands.
Methods Cell Biol. 2013;117:197-212. doi: 10.1016/B978-0-12-408143-7.00011-6.
Membrane-localized receptor proteins are involved in many signaling cascades, and diffusion and oligomerization are key processes controlling their activity. In order to study these processes in living cells, fluorescence fluctuation spectroscopy techniques have been developed that allow the quantification of concentration levels, diffusion rates, and interactions between fluorescently labeled receptor proteins at the nanomolar concentration level. This chapter presents a brief introduction to the technique and a protocol to measure and quantify the diffusion and oligomerization of human histamine1 receptor complexes in living HeLa cells using line-scanning fluorescence cross-correlation spectroscopy.
膜定位受体蛋白参与许多信号级联反应，扩散和寡聚化是控制其活性的关键过程。为了在活细胞中研究这些过程，已开发出荧光涨落光谱技术，该技术可在纳摩尔浓度水平上对荧光标记受体蛋白的浓度水平、扩散速率及其相互作用进行定量。本章简要介绍该技术，并给出一个使用线扫描荧光互相关光谱法测量和定量活HeLa细胞中人组胺1受体复合物的扩散和寡聚化的实验方案。
