Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1986 Dec 1; 103(6): 2637–2647.
PMCID: PMC2114617

Identification of an alternatively spliced site in human plasma fibronectin that mediates cell type-specific adhesion


We have compared the molecular specificities of the adhesive interactions of melanoma and fibroblastic cells with fibronectin. Several striking differences were found in the sensitivity of the two cell types to inhibition by a series of synthetic peptides modeled on the Arg-Gly-Asp-Ser (RGDS) tetrapeptide adhesion signal. Further evidence for differences between the melanoma and fibroblastic cell adhesion systems was obtained by examining adhesion to proteolytic fragments of fibronectin. Fibroblastic BHK cells spread readily on fl3, a 75-kD fragment representing the RGDS-containing, "cell-binding" domain of fibronectin, but B16-F10 melanoma cells could not. The melanoma cells were able to spread instead on f9, a 113-kD fragment derived from the large subunit of fibronectin that contains at least part of the type III connecting segment difference region (or "V" region); f7, a fragment from the small fibronectin subunit that lacks this alternatively spliced polypeptide was inactive. Monoclonal antibody and fl3 inhibition experiments confirmed the inability of the melanoma cells to use the RGDS sequence; neither molecule affected melanoma cell spreading, but both completely abrogated fibroblast adhesion. By systematic analysis of a series of six overlapping synthetic peptides spanning the entire type III connecting segment, a novel attachment site was identified in a peptide near the COOH- terminus of this region. The tetrapeptide sequence Arg-Glu-Asp-Val (REDV), which is somewhat related to RGDS, was present in this peptide in a highly hydrophilic region of the type III connecting segment. REDV appeared to be functionally important, since this synthetic tetrapeptide was inhibitory for melanoma cell adhesion to fibronectin but was inactive for fibroblastic cell adhesion. REDV therefore represents a novel adhesive recognition signal in fibronectin that possesses cell type specificity. These results suggest that, for some cell types, regulation of the adhesion-promoting activity of fibronectin may occur by alternative mRNA splicing.

Full Text

The Full Text of this article is available as a PDF (1.7M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Akiyama SK, Yamada KM. The interaction of plasma fibronectin with fibroblastic cells in suspension. J Biol Chem. 1985 Apr 10;260(7):4492–4500. [PubMed]
  • Akiyama SK, Yamada KM. Synthetic peptides competitively inhibit both direct binding to fibroblasts and functional biological assays for the purified cell-binding domain of fibronectin. J Biol Chem. 1985 Sep 5;260(19):10402–10405. [PubMed]
  • Akiyama SK, Hasegawa E, Hasegawa T, Yamada KM. The interaction of fibronectin fragments with fibroblastic cells. J Biol Chem. 1985 Oct 25;260(24):13256–13260. [PubMed]
  • Akiyama SK, Yamada SS, Yamada KM. Characterization of a 140-kD avian cell surface antigen as a fibronectin-binding molecule. J Cell Biol. 1986 Feb;102(2):442–448. [PMC free article] [PubMed]
  • Bernard MP, Kolbe M, Weil D, Chu ML. Human cellular fibronectin: comparison of the carboxyl-terminal portion with rat identifies primary structural domains separated by hypervariable regions. Biochemistry. 1985 May 21;24(11):2698–2704. [PubMed]
  • Beyth RJ, Culp LA. Complementary adhesive responses of human skin fibroblasts to the cell-binding domain of fibronectin and the heparan sulfate-binding protein, platelet factor-4. Exp Cell Res. 1984 Dec;155(2):537–548. [PubMed]
  • Boucaut JC, Darribère T, Poole TJ, Aoyama H, Yamada KM, Thiery JP. Biologically active synthetic peptides as probes of embryonic development: a competitive peptide inhibitor of fibronectin function inhibits gastrulation in amphibian embryos and neural crest cell migration in avian embryos. J Cell Biol. 1984 Nov;99(5):1822–1830. [PMC free article] [PubMed]
  • Fidler IJ. Selection of successive tumour lines for metastasis. Nat New Biol. 1973 Apr 4;242(118):148–149. [PubMed]
  • Fidler IJ. Inhibition of pulmonary metastasis by intravenous injection of specifically activated macrophages. Cancer Res. 1974 May;34(5):1074–1078. [PubMed]
  • Ginsberg M, Pierschbacher MD, Ruoslahti E, Marguerie G, Plow E. Inhibition of fibronectin binding to platelets by proteolytic fragments and synthetic peptides which support fibroblast adhesion. J Biol Chem. 1985 Apr 10;260(7):3931–3936. [PubMed]
  • Grinnell F, Feld MK. Adsorption characteristics of plasma fibronectin in relationship to biological activity. J Biomed Mater Res. 1981 May;15(3):363–381. [PubMed]
  • Hasegawa T, Hasegawa E, Chen WT, Yamada KM. Characterization of a membrane-associated glycoprotein complex implicated in cell adhesion to fibronectin. J Cell Biochem. 1985;28(4):307–318. [PubMed]
  • Haverstick DM, Cowan JF, Yamada KM, Santoro SA. Inhibition of platelet adhesion to fibronectin, fibrinogen, and von Willebrand factor substrates by a synthetic tetrapeptide derived from the cell-binding domain of fibronectin. Blood. 1985 Oct;66(4):946–952. [PubMed]
  • Hayashi M, Yamada KM. Domain structure of the carboxyl-terminal half of human plasma fibronectin. J Biol Chem. 1983 Mar 10;258(5):3332–3340. [PubMed]
  • Hopp TP, Woods KR. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. [PMC free article] [PubMed]
  • Horwitz A, Duggan K, Greggs R, Decker C, Buck C. The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin. J Cell Biol. 1985 Dec;101(6):2134–2144. [PMC free article] [PubMed]
  • Humphries MJ, Matsumoto K, White SL, Olden K. Oligosaccharide modification by swainsonine treatment inhibits pulmonary colonization by B16-F10 murine melanoma cells. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1752–1756. [PMC free article] [PubMed]
  • Humphries MJ, Olden K, Yamada KM. A synthetic peptide from fibronectin inhibits experimental metastasis of murine melanoma cells. Science. 1986 Jul 25;233(4762):467–470. [PubMed]
  • Hynes R. Molecular biology of fibronectin. Annu Rev Cell Biol. 1985;1:67–90. [PubMed]
  • Hynes RO. Fibronectins. Sci Am. 1986 Jun;254(6):42–51. [PubMed]
  • Kleinman HK, Martin GR, Fishman PH. Ganglioside inhibition of fibronectin-mediated cell adhesion to collagen. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3367–3371. [PMC free article] [PubMed]
  • Kornblihtt AR, Umezawa K, Vibe-Pedersen K, Baralle FE. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 1985 Jul;4(7):1755–1759. [PMC free article] [PubMed]
  • Kornblihtt AR, Vibe-Pedersen K, Baralle FE. Human fibronectin: cell specific alternative mRNA splicing generates polypeptide chains differing in the number of internal repeats. Nucleic Acids Res. 1984 Jul 25;12(14):5853–5868. [PMC free article] [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Laterra J, Silbert JE, Culp LA. Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin. J Cell Biol. 1983 Jan;96(1):112–123. [PMC free article] [PubMed]
  • McCarthy JB, Hagen ST, Furcht LT. Human fibronectin contains distinct adhesion- and motility-promoting domains for metastatic melanoma cells. J Cell Biol. 1986 Jan;102(1):179–188. [PMC free article] [PubMed]
  • McKeown-Longo PJ, Mosher DF. Interaction of the 70,000-mol-wt amino-terminal fragment of fibronectin with the matrix-assembly receptor of fibroblasts. J Cell Biol. 1985 Feb;100(2):364–374. [PMC free article] [PubMed]
  • Miekka SI, Ingham KC, Menache D. Rapid methods for isolation of human plasma fibronectin. Thromb Res. 1982 Jul 1;27(1):1–14. [PubMed]
  • Mosher DF. Physiology of fibronectin. Annu Rev Med. 1984;35:561–575. [PubMed]
  • Odermatt E, Tamkun JW, Hynes RO. Repeating modular structure of the fibronectin gene: relationship to protein structure and subunit variation. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6571–6575. [PMC free article] [PubMed]
  • Paul JI, Schwarzbauer JE, Tamkun JW, Hynes RO. Cell-type-specific fibronectin subunits generated by alternative splicing. J Biol Chem. 1986 Sep 15;261(26):12258–12265. [PubMed]
  • Pierschbacher MD, Ruoslahti E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984 May 3;309(5963):30–33. [PubMed]
  • Pierschbacher MD, Ruoslahti E. Variants of the cell recognition site of fibronectin that retain attachment-promoting activity. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5985–5988. [PMC free article] [PubMed]
  • Pierschbacher M, Hayman EG, Ruoslahti E. Synthetic peptide with cell attachment activity of fibronectin. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1224–1227. [PMC free article] [PubMed]
  • Plow EF, Pierschbacher MD, Ruoslahti E, Marguerie GA, Ginsberg MH. The effect of Arg-Gly-Asp-containing peptides on fibrinogen and von Willebrand factor binding to platelets. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8057–8061. [PMC free article] [PubMed]
  • Pytela R, Pierschbacher MD, Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. [PubMed]
  • Ruoslahti E, Pierschbacher MD. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. [PubMed]
  • Schwarzbauer JE, Paul JI, Hynes RO. On the origin of species of fibronectin. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1424–1428. [PMC free article] [PubMed]
  • Schwarzbauer JE, Tamkun JW, Lemischka IR, Hynes RO. Three different fibronectin mRNAs arise by alternative splicing within the coding region. Cell. 1983 Dec;35(2 Pt 1):421–431. [PubMed]
  • Silnutzer JE, Barnes DW. Effects of fibronectin-related peptides on cell spreading. In Vitro Cell Dev Biol. 1985 Jan;21(1):73–78. [PubMed]
  • Spiegel S, Yamada KM, Hom BE, Moss J, Fishman PH. Fluorescent gangliosides as probes for the retention and organization of fibronectin by ganglioside-deficient mouse cells. J Cell Biol. 1985 Mar;100(3):721–726. [PMC free article] [PubMed]
  • Spiegel S, Yamada KM, Hom BE, Moss J, Fishman PH. Fibrillar organization of fibronectin is expressed coordinately with cell surface gangliosides in a variant murine fibroblast. J Cell Biol. 1986 May;102(5):1898–1906. [PMC free article] [PubMed]
  • Tack BF, Dean J, Eilat D, Lorenz PE, Schechter AN. Tritium labeling of proteins to high specific radioactivity by reduction methylation. J Biol Chem. 1980 Sep 25;255(18):8842–8847. [PubMed]
  • Tamkun JW, Schwarzbauer JE, Hynes RO. A single rat fibronectin gene generates three different mRNAs by alternative splicing of a complex exon. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5140–5144. [PMC free article] [PubMed]
  • Thompson LK, Horowitz PM, Bentley KL, Thomas DD, Alderete JF, Klebe RJ. Localization of the ganglioside-binding site of fibronectin. J Biol Chem. 1986 Apr 15;261(11):5209–5214. [PubMed]
  • Umezawa K, Kornblihtt AR, Baralle FE. Isolation and characterization of cDNA clones for human liver fibronectin. FEBS Lett. 1985 Jul 1;186(1):31–34. [PubMed]
  • Vaheri A, Salonen EM, Vartio T. Fibronectin in formation and degradation of the pericellular matrix. Ciba Found Symp. 1985;114:111–126. [PubMed]
  • Wieland T, Birr C, Flor F. Symmetrical Boc-amino acid anhydrides for economical peptide syntheses on a solid phase. Angew Chem Int Ed Engl. 1971 May;10(5):336–336. [PubMed]
  • Woods A, Couchman JR, Johansson S, Hök M. Adhesion and cytoskeletal organisation of fibroblasts in response to fibronectin fragments. EMBO J. 1986 Apr;5(4):665–670. [PMC free article] [PubMed]
  • Yamada KM. Cell surface interactions with extracellular materials. Annu Rev Biochem. 1983;52:761–799. [PubMed]
  • Yamada KM, Kennedy DW. Dualistic nature of adhesive protein function: fibronectin and its biologically active peptide fragments can autoinhibit fibronectin function. J Cell Biol. 1984 Jul;99(1 Pt 1):29–36. [PMC free article] [PubMed]
  • Yamada KM, Kennedy DW. Amino acid sequence specificities of an adhesive recognition signal. J Cell Biochem. 1985;28(2):99–104. [PubMed]
  • Yamada KM, Critchley DR, Fishman PH, Moss J. Exogenous gangliosides enhance the interaction of fibronectin with ganglioside-deficient cells. Exp Cell Res. 1983 Feb;143(2):295–302. [PubMed]
  • Yamada KM, Kennedy DW, Grotendorst GR, Momoi T. Glycolipids: receptors for fibronectin? J Cell Physiol. 1981 Nov;109(2):343–351. [PubMed]
  • Zardi L, Carnemolla B, Balza E, Borsi L, Castellani P, Rocco M, Siri A. Elution of fibronectin proteolytic fragments from a hydroxyapatite chromatography column. A simple procedure for the purification of fibronectin domains. Eur J Biochem. 1985 Feb 1;146(3):571–579. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...