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PMID: 3305521 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Identification of an endosomal antigen specific to absorptive cells of suckling rat ileum.

The Journal of cell biology ·Vol. 105 ·No. 2 ·1987-08-00 ·Pages 691-703

Wilson JM, Whitney JA, Neutra MR

Abstract

A membrane fraction enriched in apical endosomal tubules was isolated from absorptive cells of suckling rat ileum and used as an immunogen to generate anti-endosome monoclonal antibodies. By immunofluorescence, one of these antibodies bound exclusively to the region of the apical endocytic complex in ileal absorptive cells, but not to other cell types. Immunoblot analysis showed the antigen as a diffuse 55-61-kD band which was highly enriched in the endosome fraction over whole-cell homogenate. The antigen appears to be an intramembrane glycoprotein: it partitioned primarily in the detergent phase after TX-114 extraction, and shifted to 44 kD after chemical deglycosylation. EM immunocytochemistry showed that the antibody bound to the luminal side of endosomal tubule membranes, a portion of endosomal vesicle membranes, and in endocytic pits of apical plasma membranes. However, it did not bind to multivesicular bodies, the giant lysosome, or other organelles. Immunocytochemistry after uptake with adsorbed or soluble tracer proteins showed that the antigen labeled portions of both prelysosomal pathways previously described in these cells (Gonnella, P.A., and M. R. Neutra, 1984, J. Cell Biol., 99:909-917). The function of this glycoprotein is not known, but inasmuch as it has been detected only in absorptive cells of suckling rat ileum, it may serve a function specific to these cells. Nevertheless, this endosomal antigen, designated glycoprotein (gp) 55-61, will serve as a useful marker for exploring membrane dynamics in early stages of the endocytic pathway.

MeSH Terms
Animals Antibodies, Monoclonal Antigens, Surface/analysis Cell Fractionation/methods Cell Membrane/ultrastructure Centrifugation, Density Gradient/methods Endocytosis Fluorescent Antibody Technique Ileum/ultrastructure Microscopy, Electron Rats Rats, Inbred Strains
Chemicals
Antibodies, Monoclonal Antigens, Surface
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wilson J M
Whitney J A
Neutra M R
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55 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.105.2.691
Published
1987-08-00
Pages
691-703
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114780
Subset
IM
Grants
NIADDK NIH HHS · AM-34854 · United States
NICHD NIH HHS · HD-17557 · United States
External Links
PubMed source DOI: 10.1083/jcb.105.2.691 PMC Full Text View journal details
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