G蛋白偶联受体（GPCR）的定位与转运分析。
Analysis of GPCR localization and trafficking.
作者信息
Hislop James N, von Zastrow Mark
机构信息
Department of Psychiatry, University of California, San Francisco, CA, USA.
出版信息
Methods Mol Biol. 2011;746:425-40. doi: 10.1007/978-1-61779-126-0_25.
Abstract
Localization and trafficking of G protein-coupled receptors (GPCRs) is increasingly recognized to play a fundamental role in receptor-mediated signaling and its regulation. Individual receptors, including closely homologous subtypes with otherwise similar functional properties, can differ considerably in their membrane trafficking properties. In this chapter, we describe several approaches for experimentally assessing the subcellular localization and trafficking of selected GPCRs. Firstly, we describe a flexible method for receptor localization using fluorescence microscopy. We then describe two complementary approaches, using fluorescence flow cytometry and surface biotinylation, for examining receptor internalization and trafficking in the endocytic pathway.
摘要
G蛋白偶联受体（GPCRs）的定位和运输在受体介导的信号传导及其调节中发挥着越来越重要的基础作用，这一点已得到广泛认可。包括具有相似功能特性的密切同源亚型在内的单个受体，其膜运输特性可能存在很大差异。在本章中，我们描述了几种通过实验评估所选GPCRs亚细胞定位和运输的方法。首先，我们描述一种使用荧光显微镜进行受体定位的灵活方法。然后，我们描述两种互补方法，即使用荧光流式细胞术和表面生物素化，来检测受体在内吞途径中的内化和运输。
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