A novel assay to demonstrate an intersection of the exocytic and endocytic pathways at early endosomes
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
ID 153662
Author(s) Laird, V; Spiess, M
Author(s) at UniBasel Spiess, Martin
Year 2000
Title A novel assay to demonstrate an intersection of the exocytic and endocytic pathways at early endosomes
Journal Experimental cell research
Volume 260
Number 2
Pages / Article-Number 340-5
Keywords asialoglycoprotein receptor, endocytosis, exocytosis, transferrin, basolateral sorting
Abstract The mechanism of transport of membrane proteins from the trans-Golgi to the cell surface is still poorly understood. Previous studies suggested that basolateral membrane proteins, such as the transferrin receptor and the asialoglycoprotein receptor H1, take an indirect route to the plasma membrane via an intracellular, most likely endosomal intermediate. To define this compartment we developed a biochemical assay based on the very definition of endosomes. The assay is based on internalizing anti-H1 antibodies via the endocytic cycle of the receptor itself. Internalized antibody formed immune complexes with newly synthesized H1, which had been pulse-labeled with [(35)S]sulfate and chased out of the trans-Golgi for a period of time that was insufficient for H1 to reach the surface. Hence, antibody capture occurred intracellularly. Double-immunofluorescence labeling demonstrated that antibody-containing compartments also contained transferrin and thus corresponded to early and recycling endosomes. The results therefore demonstrate an intracellular intersection of the exocytic and endocytic pathways with implications for basolateral sorting.
Publisher Elsevier
ISSN/ISBN 0014-4827
edoc-URL http://edoc.unibas.ch/dok/A5258057
Full Text on edoc No
Digital Object Identifier DOI 10.1006/excr.2000.5006
PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/11035929
ISI-Number WOS:000165205700018
Document type (ISI) Journal Article

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