Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1980 Jun 1;85(3):527–533. doi: 10.1083/jcb.85.3.527

Direct association of fibronectin and actin molecules in vitro

PMCID: PMC2111462  PMID: 6893050

Abstract

Affinity chromatography with actin-Sepharose conjugates of purified human fibronectin, normal human plasma, or serum-free culture fluid from human fibroblasts showed that fibronectin molecules can directly bind to actin. A quantitative recovery of soluble human fibronectin was accomplished by chromatography on actin immobilized on Sepharose beads. Human fibronectin molecules bound to actin-Sepharose were eluted with 0.25--0.35 M potassium bromide, and these molecules competed in a species-specific radioimmunoassay for human fibronectin. The subunits of fibronectin isolated by actin-Sepharose chromatography comigrated in SDS polyacrylamide gel electrophoresis with those of electrophoretically homogeneous fibronectin purified by conventional procedures. The efficient direct binding of fibronectin to actin suggests that interactions between these proteins might also take place in vivo but further studies are needed to elucidate the biological significance of this affinity.

Full Text

The Full Text of this article is available as a PDF (591.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ali I. U., Hynes R. O. Effects of cytochalasin B and colchicine on attachment of a major surface protein of fibroblasts. Biochim Biophys Acta. 1977 Nov 15;471(1):16–24. doi: 10.1016/0005-2736(77)90388-1. [DOI] [PubMed] [Google Scholar]
  2. Ali I. U., Mautner V., Lanza R., Hynes R. O. Restoration of normal morphology, adhesion and cytoskeleton in transformed cells by addition of a transformation-sensitive surface protein. Cell. 1977 May;11(1):115–126. doi: 10.1016/0092-8674(77)90322-1. [DOI] [PubMed] [Google Scholar]
  3. Carter W. G., Hakomori S. Isolation and partial characterization of "galactoprotein a" (LETS) and "galactoprotein b" from hamster embryo fibroblasts. Biochem Biophys Res Commun. 1976 May 23;76(2):299–308. doi: 10.1016/0006-291x(77)90725-2. [DOI] [PubMed] [Google Scholar]
  4. Chen L. B., Murray A., Segal R. A., Bushnell A., Walsh M. L. Studies on intercellular LETS glycoprotein matrices. Cell. 1978 Jun;14(2):377–391. doi: 10.1016/0092-8674(78)90123-x. [DOI] [PubMed] [Google Scholar]
  5. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  6. Dessau W., Jilek F., Adelmann B. C., Hörmann H. Similarity of antigelatin factor and cold insoluble globulin. Biochim Biophys Acta. 1978 Mar 28;533(1):227–237. doi: 10.1016/0005-2795(78)90566-4. [DOI] [PubMed] [Google Scholar]
  7. Engvall E., Ruoslahti E., Miller E. J. Affinity of fibronectin to collagens of different genetic types and to fibrinogen. J Exp Med. 1978 Jun 1;147(6):1584–1595. doi: 10.1084/jem.147.6.1584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heggeness M. H., Ash J. F., Singer S. J. Transmembrane linkage of fibronectin to intracellular actin-containing filaments in cultured human fibroblasts. Ann N Y Acad Sci. 1978 Jun 20;312:414–417. doi: 10.1111/j.1749-6632.1978.tb16822.x. [DOI] [PubMed] [Google Scholar]
  9. Hunt R. C., Brown J. C. Identification of a high molecular weight trans-membrane protein in mouse L cells. J Mol Biol. 1975 Oct 5;97(4):413–422. doi: 10.1016/s0022-2836(75)80051-9. [DOI] [PubMed] [Google Scholar]
  10. Hynes R. O. Cell surface proteins and malignant transformation. Biochim Biophys Acta. 1976 Apr 30;458(1):73–107. doi: 10.1016/0304-419x(76)90015-9. [DOI] [PubMed] [Google Scholar]
  11. Hynes R. O., Destree A. T. Relationships between fibronectin (LETS protein) and actin. Cell. 1978 Nov;15(3):875–886. doi: 10.1016/0092-8674(78)90272-6. [DOI] [PubMed] [Google Scholar]
  12. Keski-Oja J., Mosher D. F., Vaheri A. Dimeric character of fibronectin, a major cell surface-associated glycoprotein. Biochem Biophys Res Commun. 1977 Jan 24;74(2):699–706. doi: 10.1016/0006-291x(77)90359-x. [DOI] [PubMed] [Google Scholar]
  13. Keski-Oja J., Shoyab M., De Larco J. E., Todaro G. J. Rapid release of fibronectin from human lung fibroblasts by biologically active phorbol esters. Int J Cancer. 1979 Aug;24(2):218–224. doi: 10.1002/ijc.2910240214. [DOI] [PubMed] [Google Scholar]
  14. Kleinman H. K., Martin G. R., Fishman P. H. Ganglioside inhibition of fibronectin-mediated cell adhesion to collagen. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3367–3371. doi: 10.1073/pnas.76.7.3367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kleinman H. K., McGoodwin E. B., Martin G. R., Klebe R. J., Fietzek P. P., Woolley D. E. Localization of the binding site for cell attachment in the alpha1(I) chain of collagen. J Biol Chem. 1978 Aug 25;253(16):5642–5646. [PubMed] [Google Scholar]
  16. Kurkinen M., Wartiovaara J., Vaheri A. Cytochalasin B releases a major surface-associated glycoprotein, fibronectin, from cultured fibroblasts. Exp Cell Res. 1978 Jan;111(1):127–137. doi: 10.1016/0014-4827(78)90243-4. [DOI] [PubMed] [Google Scholar]
  17. Kuusela P., Ruoslahti E., Vaheri A. Polypeptides of a glycoprotein antigen (SF) present in serum and surface of normal but not of transformed chicken fibroblasts. Biochim Biophys Acta. 1975 Jan 30;379(1):295–303. doi: 10.1016/0005-2795(75)90032-x. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Mosher D. F. Cross-linking of plasma and cellular fibronectin by plasma transglutaminase. Ann N Y Acad Sci. 1978 Jun 20;312:38–42. doi: 10.1111/j.1749-6632.1978.tb16791.x. [DOI] [PubMed] [Google Scholar]
  20. Mosher D. F., Vaheri A. Thrombin stimulates the production and release of a major surface-associated glycoprotein (fibronectin) in cultures of human fibroblasts. Exp Cell Res. 1978 Mar 15;112(2):323–334. doi: 10.1016/0014-4827(78)90215-x. [DOI] [PubMed] [Google Scholar]
  21. Parsons R. G., Aldenderfer P. H., Kowal R. Detection of a human serum DNA-binding protein associated with malignant disease. J Natl Cancer Inst. 1979 Jul;63(1):43–47. [PubMed] [Google Scholar]
  22. Ruoslahti E., Vaheri A. Interaction of soluble fibroblast surface antigen with fribrinogen and fibrin. J Exp Med. 1975 Feb 1;141(2):497–501. doi: 10.1084/jem.141.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Saba T. M., Blumenstock F. A., Weber P., Kaplan J. E. Physiologic role of cold-insoluble globulin in systemic host defense: implications of its characterization as the opsonic alpha 2-surface-binding glycoprotein. Ann N Y Acad Sci. 1978 Jun 20;312:43–55. doi: 10.1111/j.1749-6632.1978.tb16792.x. [DOI] [PubMed] [Google Scholar]
  24. Sen A., Todaro G. J. A murine sarcoma virus-associated protein kinase: interaction with actin and microtubular protein. Cell. 1979 Jun;17(2):347–356. doi: 10.1016/0092-8674(79)90161-2. [DOI] [PubMed] [Google Scholar]
  25. Singer I. I. The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts. Cell. 1979 Mar;16(3):675–685. doi: 10.1016/0092-8674(79)90040-0. [DOI] [PubMed] [Google Scholar]
  26. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  27. Vaheri A., Kurkinen M., Lehto V. P., Linder E., Timpl R. Codistribution of pericellular matrix proteins in cultured fibroblasts and loss in transformation: fibronectin and procollagen. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4944–4948. doi: 10.1073/pnas.75.10.4944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vaheri A., Mosher D. F. High molecular weight, cell surface-associated glycoprotein (fibronectin) lost in malignant transformation. Biochim Biophys Acta. 1978 Sep 18;516(1):1–25. doi: 10.1016/0304-419x(78)90002-1. [DOI] [PubMed] [Google Scholar]
  29. Willingham M. C., Yamada K. M., Yamada S. S., Pouysségur J., Pastan I. Microfilament bundles and cell shape are related to adhesiveness to substratum and are dissociable from growth control in cultured fibroblasts. Cell. 1977 Mar;10(3):375–380. doi: 10.1016/0092-8674(77)90024-1. [DOI] [PubMed] [Google Scholar]
  30. Yamada K. M., Olden K. Fibronectins--adhesive glycoproteins of cell surface and blood. Nature. 1978 Sep 21;275(5677):179–184. doi: 10.1038/275179a0. [DOI] [PubMed] [Google Scholar]
  31. Yamada K. M., Weston J. A. Isolation of a major cell surface glycoprotein from fibroblasts. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3492–3496. doi: 10.1073/pnas.71.9.3492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zardi L., Siri A., Carnemolla B., Santi L., Gardner W. D., Hoch S. O. Fibronectin: a chromatin-associated protein? Cell. 1979 Nov;18(3):649–657. doi: 10.1016/0092-8674(79)90120-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES