A cell-free assay for endocytosis of E-cadherin.

E-Cadherin is a key cell-cell adhesion molecule at adherens junctions (AJs) and undergoes endocytosis when AJs are disrupted by the action of extracellular signals. To elucidate the mechanism of this endocytosis, we developed a new cell-free assay system for this reaction using the AJ-enriched fraction from rat liver. Non-trans-interacting E-cadherin, but not the trans-interacting one, underwent endocytosis in a clathrin-dependent manner. The endocytosis of trans-interacting E-cadherin was inhibited by Rac and Cdc42 small G proteins, which were activated by trans-interacting E-cadherin. Here, we describe the assay method for the endocytosis of E-cadherin from the AJ-enriched fraction prepared from rat liver.