Reverse endocytosis of transmembrane ephrin-B ligands via a clathrin-mediated pathway.

Parker M, Roberts R, Enriquez M, Zhao X, Takahashi T, Pat Cerretti D, Daniel T, Chen J
Biochem Biophys Res Commun. 2004 323 (1): 17-23

PMID: 15351694 · DOI:10.1016/j.bbrc.2004.07.209

Eph/ephrin receptors and ligands mediate cell-cell interaction through reciprocal signaling upon juxtacrine contact, and play a critical role in embryonic patterning, neuronal targeting, and vascular assembly. To study transmembrane ephrin-B ligand trafficking, we determined the cellular localization of ephrin-B1-GFP upon engagement by EphB1. Under normal culture conditions ephrin-B1-GFP is localized to the plasma membrane, mostly at the lateral cell borders. Addition of soluble EphB1-Fc receptor induces ephrin-B1-GFP clustering on the cell surface and subsequent internalization, as judged by biochemical studies, electron microscopy, and co-localization with endosomal markers. A dominant-negative mutant of dynamin or potassium depletion blocks ephrin-B1 endocytosis. These results suggest that ephrin-B1 internalization is an active receptor-mediated process that utilizes the clathrin-mediated endocytic pathway.

Copyright 2004 Elsevier Inc.

MeSH Terms (27)

Animals Biotinylation Blotting, Western Cell Membrane Cells, Cultured Cell Separation CHO Cells Clathrin Cricetinae DNA, Complementary Dynamins Endocytosis Ephrin-B1 Flow Cytometry Genes, Dominant Genetic Vectors Humans Ligands Microscopy, Confocal Microscopy, Electron Mutation Potassium Receptors, Eph Family Signal Transduction Time Factors Transfection Umbilical Veins

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