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PMID: 2687287 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Polarized endocytosis by Madin-Darby canine kidney cells transfected with functional chicken liver glycoprotein receptor.

The Journal of cell biology ·Vol. 109 ·No. 6 Pt 1 ·1989-12-00 ·Pages 2809-16

Graeve L, Drickamer K, Rodriguez-Boulan E

Abstract

We have studied the expression of the chicken hepatic glycoprotein receptor (chicken hepatic lectin [CHL]) in Madin-Darby canine kidney (MDCK) cells, by transfection of its cDNA under the control of a retroviral promotor. Transfected cell lines stably express 87,000 surface receptors/cell with a kd = 13 nM. In confluent monolayers, approximately 40% of CHL is localized at the plasma membrane. 98% of the surface CHL is expressed at the basolateral surface where it performs polarized endocytosis and degradation of glycoproteins carrying terminal N-acetylglucosamine at a rate of 50,000 ligand molecules/h. Studies of the half-life of metabolically labeled receptor and of the stability of biotinylated cell surface receptor after internalization indicate that transfected CHL performs several rounds of uptake and recycling before it gets degraded. The successful expression of a functional basolateral receptor in MDCK cells opens the way for the characterization of the mechanisms that control targeting and recycling of proteins to the basolateral membrane of epithelial cells.

MeSH Terms
Animals Biotin Cell Line Chickens Cloning, Molecular DNA/genetics Dogs Endocytosis Fluorescent Antibody Technique Glycoproteins/metabolism Immunoblotting Kidney Kinetics Liver/immunology Platelet Glycoprotein GPIb-IX Complex Platelet Membrane Glycoproteins Receptors, Immunologic/genetics,metabolism Transfection
Chemicals
Glycoproteins Platelet Glycoprotein GPIb-IX Complex Platelet Membrane Glycoproteins Receptors, Immunologic glycoprotein receptor GPIb-IX Biotin DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Graeve L
Department of Cell Biology and Anatomy, Cornell University Medical College, New York 10021.
Drickamer K
Rodriguez-Boulan E
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.109.6.2809
Published
1989-12-00
Pages
2809-16
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115943
Subset
IM
Grants
NCRR NIH HHS · 507 RR05 396 · United States
NIGMS NIH HHS · GM-34107 · United States
External Links
PubMed source DOI: 10.1083/jcb.109.6.2809 PMC Full Text View journal details
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