Li Chunying, Naren Anjaparavanda P
Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Methods Mol Biol. 2011;741:255-70. doi: 10.1007/978-1-61779-117-8_17.
Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel localized primarily at the apical surface of epithelial cells lining the airway, gut, exocrine glands, etc., where it is responsible for transepithelial salt and water transport. A growing number of proteins have been reported to interact directly or indirectly with CFTR chloride channel, suggesting that CFTR might regulate the activities of other ion channels, receptors, and transporters, in addition to its role as a chloride conductor. Most interactions occur primarily between the opposing terminal tails (N or C) of CFTR and its binding partners, either directly or mediated through various PDZ domain-containing proteins. This chapter describes methods we developed to cross-link CFTR into a macromolecular complex to identify and analyze the assembly and regulation of CFTR-containing complexes in the plasma membrane.
囊性纤维化跨膜传导调节因子(CFTR)是一种氯离子通道,主要定位于气道、肠道、外分泌腺等上皮细胞的顶端表面,在这些部位负责跨上皮盐和水的运输。据报道,越来越多的蛋白质可直接或间接与CFTR氯离子通道相互作用,这表明CFTR除了作为氯离子导体的作用外,可能还调节其他离子通道、受体和转运蛋白的活性。大多数相互作用主要发生在CFTR相对的末端尾巴(N端或C端)与其结合伙伴之间,要么直接发生,要么通过各种含PDZ结构域的蛋白质介导。本章介绍了我们开发的将CFTR交联成大分子复合物的方法,以识别和分析质膜中含CFTR复合物的组装和调节。