DOI: 10.1083/jcb.149.2.521. Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis

Defining Fibronectin's Cell Adhesion Synergy Site by Site-Directed Mutagenesis

10.1083/jcb.149.2.521

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

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.

Bibliography

Redick, S. D., Settles, D. L., Briscoe, G., & Erickson, H. P. (2000). Defining Fibronectinâs Cell Adhesion Synergy Site by Site-Directed Mutagenesis. The Journal of Cell Biology, 149(2), 521â527.

Authors 4

Sambra D. Redick (first)
Daniel L. Settles (additional)
Gina Briscoe (additional)
Harold P. Erickson (additional)

References 28 Referenced 157

10.1016/S0021-9258(17)31456-4 / J. Biol. Chem. / The short amino acid sequence Pro-His-Ser-Arg-Asn in human fibronectin enhances cell-adhesive function by Aota (1994)
10.1016/S0021-9258(18)53809-6 / J. Biol. Chem. / Cell- and heparin-binding domains of the hexabrachion arm identified by tenascin expression proteins by Aukhil (1993)
10.1074/jbc.273.38.24744 / J. Biol. Chem. / The cation-binding domain from the alpha subunit of integrin α5β1 is a minimal domain for fibronectin recognition by Banères (1998)
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10.1016/S0021-9258(17)34137-6 / J. Biol. Chem. / Identification of a novel integrin binding site in fibronectin. Differential utilization by β3 integrins by Bowditch (1994)
10.1042/bj3440527 / Biochem. J. / Fine mapping of inhibitory anti-α5 monoclonal antibody epitopes that differentially affect integrin-ligand binding by Burrows (1999)
10.1016/S0021-9258(18)89656-9 / J. Biol. Chem. / Structure of human platelet membrane glycoproteins IIb and IIIa as determined by electron microscopy by Carrell (1985)
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10.1074/jbc.270.37.21612 / J. Biol. Chem. / Requirement for the synergy site for cell adhesion to fibronectin depends on the activation state of integrin α5β1 by Danen (1995)
10.1074/jbc.273.52.34710 / J. Biol. Chem. / Two-stage activation for α5β1 integrin binding to surface-adsorbed fibronectin by Garcia (1999)
10.1074/jbc.272.10.6159 / J. Biol. Chem. / Structural requirements for biological activity of the ninth and tenth FIII domains of human fibronectin by Grant (1997)
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{'key': '2023072905314783900_McClayandHertzler1999', 'first-page': '9', 'article-title': 'Quantitative measurement of cell adhesiveness', 'volume-title': 'Current Protocols in Cell Biology', 'author': 'McClay', 'year': '1999'} / Current Protocols in Cell Biology / Quantitative measurement of cell adhesiveness by McClay (1999)
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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 12:45 p.m.)
Deposited	2 years, 1 month ago (July 29, 2023, 1:32 a.m.)
Indexed	1 month ago (July 28, 2025, 2:29 a.m.)
Issued	25 years, 4 months ago (April 17, 2000)
Published	25 years, 4 months ago (April 17, 2000)
Published Online	25 years, 4 months ago (April 17, 2000)
Published Print	25 years, 4 months ago (April 17, 2000)

Funders 0

None

BibTeX

@article{Redick_2000, title={Defining Fibronectin’s Cell Adhesion Synergy Site by Site-Directed Mutagenesis}, volume={149}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.149.2.521}, DOI={10.1083/jcb.149.2.521}, number={2}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Redick, Sambra D. and Settles, Daniel L. and Briscoe, Gina and Erickson, Harold P.}, year={2000}, month=apr, pages={521–527} }

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    "date-parts": [
      [
        2000,
        4,
        17
      ]
    ]
  },
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  "journal-issue": {
    "issue": "2",
    "published-print": {
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          17
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  "URL": "http://dx.doi.org/10.1083/jcb.149.2.521",
  "relation": {},
  "ISSN": [
    "0021-9525",
    "1540-8140"
  ],
  "subject": [],
  "published": {
    "date-parts": [
      [
        2000,
        4,
        17
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}