Wang Yingqun
Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, Philadelphia, PA, USA.
Methods Mol Biol. 2012;839:27-41. doi: 10.1007/978-1-61779-510-7_3.
The planar cell polarity (PCP) pathway is a β-catenin-independent branch of the Wnt signaling cascade. In vertebrate embryos PCP signaling regulates morphogenetic events including convergent extension (CE) movements during gastrualtion. Xenopus embryo has been established as an excellent model system to dissect PCP signaling in vertebrates because morphogenetic cell behaviors including CE can easily be monitored in vivo. Xenopus Paraxial protocadherin (xPAPC) is a transmembrane protein which serves as a link between patterning factors in the Spemann's organizer and regulators of the morphogenetic movements. xPAPC regulates morphogenesis in part by modulating cell adhesion and PCP signaling. Here two methods, GST pull-down assay and yeast two-hybrid assay, are described for the identification of xPAPC interacting proteins to elucidate the mechanism by which xPAPC regulates PCP signaling.
平面细胞极性（PCP）通路是Wnt信号级联中不依赖β-连环蛋白的一个分支。在脊椎动物胚胎中，PCP信号传导调节形态发生事件，包括原肠胚形成期间的汇聚延伸（CE）运动。非洲爪蟾胚胎已被确立为研究脊椎动物PCP信号传导的优秀模型系统，因为包括CE在内的形态发生细胞行为可以在体内轻松监测。非洲爪蟾近轴原钙黏蛋白（xPAPC）是一种跨膜蛋白，它是斯佩曼组织者中的模式形成因子与形态发生运动调节因子之间的联系纽带。xPAPC部分通过调节细胞黏附和PCP信号传导来调节形态发生。本文描述了两种方法，即GST下拉分析和酵母双杂交分析，用于鉴定与xPAPC相互作用的蛋白，以阐明xPAPC调节PCP信号传导的机制。
