Identification of the peptide sequences within the EIIIA (EDA) segment of fibronectin that mediate integrin α9β1-dependent cellular activities

Arti V. Shinde, Christopher Bystroff, Chunyu Wang, Mariette G. Vogelezang, Peter A. Vincent, Richard O. Hynes, Livingston Van De Water

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

Alternative splicing of the fibronectin (FN) gene transcript provides an efficient mechanism for generating functionally appropriate forms of this adhesive glycoprotein in situ. Cellular FNs that include the EIIIA and/or EIIIB FN-III segments are prominently expressed during embryogenesis, wound healing, tumor progression, and inflammation. However, the roles of this domain in altering overall FN protein structure and regulating cellular function remain unclear. We previously reported that two integrins, α9β1 and α4β1, ligate the EIIIA segment (Liao, Y. F., Gotwals, P. J., Koteliansky, V. E., Sheppard, D., and Van De Water, L. (2002) J. Biol. Chem. 277, 14467-14474) and that the epitopes for function-blocking monoclonal antibodies lie within the C-C′ loop of EIIIA (Liao, Y. F., Wieder, K. G., Classen, J. M., and Van De Water, L. (1999) J. Biol. Chem. 274, 17876-17884). We have now performed site-directed mutagenesis within the EIIIA segment and carried out cell adhesion assays on the semutant EIIIAs. We find that the Asp41 and Gly42 residues within the C-C′ loop of EIIIA are necessary for integrin α9β1 binding. Synthetic peptides based on the predicted important amino acid sequence from the C-C′ loop encode sufficient information to completely inhibit α9β1-mediated cell adhesion. We also report that EIIIA promotes filopodial formation in α9β1-expressing cells accompanied by Cdc42 activation. Our data provide a cellular activity for the EIIIA segment, evidence for conformational lability, and peptide sequences for probing EIIIA functions in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)2858-2870
Number of pages13
JournalJournal of Biological Chemistry
Volume283
Issue number5
DOIs
StatePublished - Feb 1 2008
Externally publishedYes

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Fibronectins
Integrins
Cell adhesion
Cell Adhesion
Peptides
Mutagenesis
Blocking Antibodies
Water
Alternative Splicing
Cellular Structures
Site-Directed Mutagenesis
Adhesives
Wound Healing
Embryonic Development
Tumors
Epitopes
Amino Acid Sequence
Assays
Glycoproteins
Genes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Identification of the peptide sequences within the EIIIA (EDA) segment of fibronectin that mediate integrin α9β1-dependent cellular activities. / Shinde, Arti V.; Bystroff, Christopher; Wang, Chunyu; Vogelezang, Mariette G.; Vincent, Peter A.; Hynes, Richard O.; Van De Water, Livingston.

In: Journal of Biological Chemistry, Vol. 283, No. 5, 01.02.2008, p. 2858-2870.

Research output: Contribution to journalArticle

Shinde, Arti V. ; Bystroff, Christopher ; Wang, Chunyu ; Vogelezang, Mariette G. ; Vincent, Peter A. ; Hynes, Richard O. ; Van De Water, Livingston. / Identification of the peptide sequences within the EIIIA (EDA) segment of fibronectin that mediate integrin α9β1-dependent cellular activities. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 5. pp. 2858-2870.
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AU - Shinde, Arti V.

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AU - Vogelezang, Mariette G.

AU - Vincent, Peter A.

AU - Hynes, Richard O.

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