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.
Sakisaka
Toshiaki
T
Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine, Suita, Japan.
Takai
Yoshimi
Y
D022005:Adherens Junctions / Q000378:metabolism; D000818:Animals; D001681:Biological Assay; D015820:Cadherins / Q000378:metabolism*; D002474:Cell-Free System; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D022161:Transport Vesicles / Q000201:enzymology / Q000378:metabolism*; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D020744:rac GTP-Binding Proteins / Q000378:metabolism
D015820:Cadherins; D002966:Clathrin; D010446:Peptide Fragments; D011994:Recombinant Proteins; D020764:cdc42 GTP-Binding Protein; D020744:rac GTP-Binding Proteins