Study of the transit of an integral membrane protein from secretory granules through the plasma membrane of secreting rat basophilic leukemia cells using a specific monoclonal antibody.

The Journal of cell biology ·Vol. 102 ·No. 2 ·1986-02-00 ·Pages 516-22

Bonifacino JS, Perez P, Klausner RD, Sandoval IV

Abstract

The monoclonal antibody 5G10 reacted specifically with an 80-kD integral membrane protein in rat basophilic leukemia (RBL) cells. Immunofluorescence microscopy studies of RBL cells, fixed and permeabilized, revealed that the 80-kD protein was located in the membrane of cytoplasmic vesicles. The vesicles were identified as secretory granules by their content in immunoreactive serotonin. Expression of the 5G10 antigen on the surface of unstimulated RBL cells was low. However, RBL cells stimulated to secrete with anti-dinitrophenyl IgE followed by dinitrophenyl-bovine serum albumin or with the Ca2+ ionophore A-23187 displayed an increased expression of the antigen on their surface. Surface exposure of the 5G10 antigen was maximal at 5 min after stimulation of secretion. Removal of dinitrophenyl-bovine serum albumin from the incubation medium resulted in internalization of 50% of the antigen within 10 min.

MeSH Terms

Animals Antibodies, Monoclonal Basophils/metabolism Cell Compartmentation Cell Line Cell Membrane/metabolism Cytoplasmic Granules/metabolism Endocytosis Fluorescent Antibody Technique Intracellular Membranes/metabolism Leukemia/metabolism Membrane Proteins/immunology,metabolism Molecular Weight Rats

Chemicals

Antibodies, Monoclonal Membrane Proteins

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Bonifacino J S

Perez P

Klausner R D

Sandoval I V

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24 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.102.2.516

Published

1986-02-00

Pages

516-22

Language

English

Region

United States

NLM ID

0375356

PMCID

PMC2114069

Subset

External Links

PubMed source DOI: 10.1083/jcb.102.2.516 PMC Full Text View journal details

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Study of the transit of an integral membrane protein from secretory granules through the plasma membrane of secreting rat basophilic leukemia cells using a specific monoclonal antibody.

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