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Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis
Fibronectin's RGD-mediated binding to the α5β1 integrin is dramatically enhanced by a synergy site within fibronectin III domain 9 (FN9). Guided by the crystal structure of the cell-binding domain, we selected amino acids in FN9 that project in the same direction as the RGD, presumably toward t...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2000
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2175162/ https://www.ncbi.nlm.nih.gov/pubmed/10769040 |
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author | Redick, Sambra D. Settles, Daniel L. Briscoe, Gina Erickson, Harold P. |
author_facet | Redick, Sambra D. Settles, Daniel L. Briscoe, Gina Erickson, Harold P. |
author_sort | Redick, Sambra D. |
collection | PubMed |
description | Fibronectin's RGD-mediated binding to the α5β1 integrin is dramatically enhanced by a synergy site within fibronectin III domain 9 (FN9). Guided by the crystal structure of the cell-binding domain, we selected amino acids in FN9 that project in the same direction as the RGD, presumably toward the integrin, and mutated them to alanine. R1379 in the peptide PHSRN, and the nearby R1374 have been shown previously to be important for α5β1-mediated adhesion (Aota, S., M. Nomizu, and K.M. Yamada. 1994. J. Biol. Chem. 269:24756–24761). Our more extensive set of mutants showed that R1379 is the key residue in the synergistic effect, but other residues contribute substantially. R1374A decreased adhesion slightly by itself, but the double mutant R1374A-R1379A was significantly less adhesive than R1379A alone. Single mutations of R1369A, R1371A, T1385A, and N1386A had negligible effects on cell adhesion, but combining these substitutions either with R1379A or each other gave a more dramatic reduction of cell adhesion. The triple mutant R1374A/P1376A/R1379A had no detectable adhesion activity. We conclude that, in addition to the R of the PHRSN peptide, other residues on the same face of FN9 are required for the full synergistic effect. The integrin-binding synergy site is a much more extensive surface than the small linear peptide sequence. |
format | Text |
id | pubmed-2175162 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2000 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21751622008-05-01 Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis Redick, Sambra D. Settles, Daniel L. Briscoe, Gina Erickson, Harold P. J Cell Biol Original Article Fibronectin's RGD-mediated binding to the α5β1 integrin is dramatically enhanced by a synergy site within fibronectin III domain 9 (FN9). Guided by the crystal structure of the cell-binding domain, we selected amino acids in FN9 that project in the same direction as the RGD, presumably toward the integrin, and mutated them to alanine. R1379 in the peptide PHSRN, and the nearby R1374 have been shown previously to be important for α5β1-mediated adhesion (Aota, S., M. Nomizu, and K.M. Yamada. 1994. J. Biol. Chem. 269:24756–24761). Our more extensive set of mutants showed that R1379 is the key residue in the synergistic effect, but other residues contribute substantially. R1374A decreased adhesion slightly by itself, but the double mutant R1374A-R1379A was significantly less adhesive than R1379A alone. Single mutations of R1369A, R1371A, T1385A, and N1386A had negligible effects on cell adhesion, but combining these substitutions either with R1379A or each other gave a more dramatic reduction of cell adhesion. The triple mutant R1374A/P1376A/R1379A had no detectable adhesion activity. We conclude that, in addition to the R of the PHRSN peptide, other residues on the same face of FN9 are required for the full synergistic effect. The integrin-binding synergy site is a much more extensive surface than the small linear peptide sequence. The Rockefeller University Press 2000-04-17 /pmc/articles/PMC2175162/ /pubmed/10769040 Text en © 2000 The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Original Article Redick, Sambra D. Settles, Daniel L. Briscoe, Gina Erickson, Harold P. Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis |
title | Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis |
title_full | Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis |
title_fullStr | Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis |
title_full_unstemmed | Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis |
title_short | Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis |
title_sort | defining fibronectin's cell adhesion synergy site by site-directed mutagenesis |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2175162/ https://www.ncbi.nlm.nih.gov/pubmed/10769040 |
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