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PMID: 2380248 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Inhibition of binding of fibronectin to matrix assembly sites by anti-integrin (alpha 5 beta 1) antibodies.

The Journal of cell biology ·Vol. 111 ·No. 2 ·1990-08-00 ·Pages 699-708

Fogerty FJ, Akiyama SK, Yamada KM, Mosher DF

Abstract

Fibroblasts have cell surface sites that mediate assembly of plasma and cellular fibronectin into the extracellular matrix. Cell adhesion to fibronectin can be mediated by the interaction of an integrin (alpha 5 beta 1) with the Arg-Gly-Asp-Ser (RGDS)-containing cell adhesion region of fibronectin. We have attempted to elucidate the role of the alpha 5 beta 1 fibronectin receptor in assembly of fibronectin in matrices. Rat monoclonal antibody mAb 13, which recognizes the integrin beta 1 subunit, completely blocked binding and matrix assembly of 125I-fibronectin as well as binding of the 125I-70-kD amino-terminal fragment of fibronectin (70 kD) to fibroblast cell layers. Fab fragments of the anti-beta 1 antibody were also inhibitory. Antibody mAb 16, which recognizes the integrin alpha 5 subunit, partially blocked binding of 125I-fibronectin and 125I-70-kD. When cell layers were coincubated with fluoresceinated fibronectin and either anti-beta 1 or anti-alpha 5, anti-beta 1 was a more effective inhibitor than anti-alpha 5 of binding of labeled fibronectin to the cell layer. Inhibition of 125I-fibronectin binding by anti-beta 1 IgG occurred within 20 min. Inhibition of 125I-fibronectin binding by anti-beta 1 Fab fragments or IgG could not be overcome with increasing concentrations of fibronectin, suggesting that anti-beta 1 and exogenous fibronectin may not compete for the same binding site. No beta 1-containing integrin bound to immobilized 70 kD. These data indicate that the beta 1 subunit plays an important role in binding and assembly of exogenous fibronectin, perhaps by participation in the organization, regeneration, or cycling of the assembly site rather than by a direct interaction with fibronectin.

MeSH Terms
Amino Acid Sequence Antibodies, Monoclonal Antigen-Antibody Complex Cell Line Cells, Cultured Chromatography, Affinity Electrophoresis, Polyacrylamide Gel Extracellular Matrix/metabolism Fibroblasts/cytology Fibronectins/antagonists & inhibitors,metabolism Humans Immunoglobulin Fab Fragments Integrins/immunology,isolation & purification,metabolism Kinetics Membrane Proteins/isolation & purification,metabolism Molecular Sequence Data Osteosarcoma Skin/cytology Tumor Cells, Cultured/cytology
Chemicals
Antibodies, Monoclonal Antigen-Antibody Complex Fibronectins Immunoglobulin Fab Fragments Integrins Membrane Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Fogerty F J
Department of Medicine, University of Wisconsin, Madison 53706.
Akiyama S K
Yamada K M
Mosher D F
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.111.2.699
Published
1990-08-00
Pages
699-708
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2116209
Subset
IM
Grants
NICHD NIH HHS · HD07118 · United States
NHLBI NIH HHS · HL21644 · United States
External Links
PubMed source DOI: 10.1083/jcb.111.2.699 PMC Full Text View journal details
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