Choi Wangsun, Goldfarb Dennis, Yan Feng, Major Michael B, Fanning Alan S, Peifer Mark
Department of Biology, University of North Carolina at Chapel Hill, CB#3280, Chapel Hill, NC 27599-3280, USA.
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO, USA63110.
Biol Open. 2025 Feb 15;14(2). doi: 10.1242/bio.061811. Epub 2025 Jan 30.
The network of proteins at the interface between cell-cell adherens junctions and the actomyosin cytoskeleton provides robust yet dynamic connections that facilitate cell shape change and motility. While this was initially thought to be a simple linear connection via classic cadherins and their associated catenins, we now have come to appreciate that many more proteins are involved, providing robustness and mechanosensitivity. Defining the full set of proteins in this network remains a key objective in our field. Proximity proteomics provides a means to define these networks. Mammalian Afadin and its Drosophila homolog Canoe are key parts of this protein network, facilitating diverse cell shape changes during gastrulation and other events of embryonic morphogenesis. Here we report results of several proximity proteomics screens, defining proteins in the neighborhood of both the N- and C-termini of mammalian Afadin in the premier epithelial model, MDCK cells. We compare our results with previous screens done in other cell types, and with proximity proteomics efforts with other junctional proteins. These reveal the value of multiple screens in defining the full network of neighbors and offer interesting insights into the overlap in protein composition between different epithelial cell junctions.
细胞间黏着连接与肌动球蛋白细胞骨架之间界面处的蛋白质网络提供了强大而动态的连接，促进细胞形状变化和运动。虽然最初认为这是通过经典钙黏着蛋白及其相关连环蛋白形成的简单线性连接，但我们现在认识到涉及更多蛋白质，从而提供了稳健性和机械敏感性。确定该网络中的全套蛋白质仍然是我们领域的一个关键目标。邻近蛋白质组学提供了一种定义这些网络的方法。哺乳动物的Afadin及其果蝇同源物Canoe是该蛋白质网络的关键部分，在原肠胚形成和胚胎形态发生的其他事件中促进多种细胞形状变化。在这里，我们报告了几项邻近蛋白质组学筛选的结果，确定了在主要上皮模型MDCK细胞中哺乳动物Afadin的N端和C端附近的蛋白质。我们将我们的结果与之前在其他细胞类型中进行的筛选结果以及对其他连接蛋白的邻近蛋白质组学研究结果进行了比较。这些结果揭示了多次筛选在定义完整的邻近网络中的价值，并为不同上皮细胞连接之间蛋白质组成的重叠提供了有趣的见解。
