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PMID: 1560034 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A synthetic peptide derived from the carboxy terminus of the laminin A chain represents a binding site for the alpha 3 beta 1 integrin.

The Journal of cell biology ·Vol. 117 ·No. 2 ·1992-04-00 ·Pages 449-59

Gehlsen KR, Sriramarao P, Furcht LT, Skubitz AP

Abstract

The purpose of this study was to identify the binding site(s) within laminin for the alpha 3 beta 1 integrin receptor. It has been previously shown, using proteolytic fragments and anti-laminin antibodies, that the region in laminin for alpha 3 beta 1 integrin binding is localized to the carboxy-terminal region at the end of the long arm (Gehlsen, K. R., E. Engvall, K. Dickerson, W. S. Argraves, and E. Ruoslahti. 1989. J. Biol. Chem. 264:19034-19038; Tomaselli, K. J., D. E. Hall, L. T. Reichardt, L. A. Flier, K. R. Gehlsen, D. C. Turner, and S. Carbonetto. 1990. Neuron. 5:651-662). Using synthetic peptides, we have identified an amino acid sequence within the carboxy-terminal region of the laminin A chain that is recognized by the alpha 3 beta 1 integrin. The amino acid sequence represented by the synthetic peptide GD-6 (KQNCLSSRASFRGCVRNLRLSR residues numbered 3011 to 3032) of the globular domain of the murine A chain supports cell attachment and inhibits cell adhesion to laminin-coated surfaces. By affinity chromatography, peptide GD-6-Sepharose specifically bound solubilized alpha 3 beta 1 from extracts of surface-iodinated cells in a cation-dependent manner, while it did not bind other integrins. In addition, exogenous peptide GD-6 specifically eluted bound alpha 3 beta 1 from laminin-Sepharose columns but did not elute the alpha 3 beta 1 integrin from a fibronectin-Sepharose column. Using integrin subunit-specific monoclonal antibodies, only those antibodies against the alpha 3 and beta 1 subunits inhibited cell adhesion to peptide GD-6-coated surfaces. Finally, a polyclonal antibody made against peptide GD-6 reacted specifically with both murine and human laminin and significantly inhibited cell adhesion to laminin-coated surfaces but not those coated with other matrix proteins. These results identify the laminin A chain amino acid sequence of peptide GD-6 as representing a binding site in laminin for the alpha 3 beta 1 integrin.

MeSH Terms
Amino Acid Sequence Binding Sites Cell Adhesion Chromatography, Affinity Humans Integrin alpha3beta1 Integrins/immunology,isolation & purification,metabolism Laminin/chemistry,metabolism,pharmacology Molecular Sequence Data Peptide Fragments/chemistry,metabolism Peptides/chemistry,metabolism Tumor Cells, Cultured
Chemicals
Integrin alpha3beta1 Integrins Laminin Peptide Fragments Peptides GD-6 peptide
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Gehlsen K R
California Institute for Biological Research, La Jolla 92037.
Sriramarao P
Furcht L T
Skubitz A P
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49 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.117.2.449
Published
1992-04-00
Pages
449-59
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289418
Subset
IM
Grants
NCI NIH HHS · CA29995 · United States
NCI NIH HHS · CA57202 · United States
External Links
PubMed source DOI: 10.1083/jcb.117.2.449 PMC Full Text View journal details
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