Mapping transmembrane binding partners for E-cadherin ectodomains

Abstract

We combine proximity labeling and single molecule binding assays, to discover novel transmembrane protein interactions in cells. We first screen for proximal membrane proteins by tagging the extracellular and cytoplasmic regions of the protein of interest with TurboID biotin ligase, and then test direct binding interactions using single molecule Atomic Force Microscopy.Using this approach, we identify previously unknown direct interactions between membrane proteins and the extracellular regions of cadherin adhesion proteins in epithelial cells.