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PMID: 1935897 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro binding of the asialoglycoprotein receptor to the beta adaptin of plasma membrane coated vesicles.

The EMBO journal ·Vol. 10 ·No. 12 ·1991-12-00 ·Pages 3735-42

Beltzer JP, Spiess M

Abstract

The asialoglycoprotein (ASGP) receptor was used to probe total clathrin-coated vesicle proteins and purified adaptor proteins (APs) which had been fractionated by gel electrophoresis and transferred to nitrocellulose. The receptor was found to interact with proteins of approximately 100 kDa. The cytoplasmic domain of the ASGP receptor subunit H1 fused to dihydrofolate reductase competed for receptor binding to the 100 kDa polypeptide in the plasma membrane-type AP complexes (AP-2). A fusion protein containing the cytoplasmic domain of the endocytic mutant haemagglutinin HA-Y543 also competed, but a protein with the wild-type haemagglutinin sequence did not. This indicates that the observed interaction is specific for the cytoplasmic domain of the receptor and involves the tyrosine signal for endocytosis. When fractionated by gel electrophoresis in the presence of urea, the ASGP receptor binding polypeptide displayed a characteristic shift in electrophoretic mobility identifying it as the beta adaptin. Partial proteolysis of the AP-2 preparation followed by the receptor binding assay revealed that the aminoterminal domain of the beta adaptin contains the binding site for receptors.

MeSH Terms
Adaptor Protein Complex beta Subunits Amino Acid Sequence Animals Asialoglycoprotein Receptor Asialoglycoproteins/metabolism Blotting, Western Cattle Cell Membrane/metabolism Coated Pits, Cell-Membrane/metabolism Electrophoresis, Polyacrylamide Gel Endocytosis Humans Hydrolysis Molecular Sequence Data Proteins/metabolism Receptors, Immunologic/metabolism
Chemicals
Adaptor Protein Complex beta Subunits Asialoglycoprotein Receptor Asialoglycoproteins Proteins Receptors, Immunologic
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Beltzer J P
Department of Biochemistry, University of Basel, Switzerland.
Spiess M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1991-12-00
Pages
3735-42
Language
English
Region
England
NLM ID
8208664
PMCID
PMC453108
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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