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. 1978 Jun 1;147(6):1779–1791. doi: 10.1084/jem.147.6.1779

Synthesis of fibronectin by cultured human endothelial cells

PMCID: PMC2184307  PMID: 355597

Abstract

Plasma fibronectin is probably the major nonimmune particulate opsonin in blood and is cross-linked to fibrin during the final stage of blood coagulation. Fibronectin also occurs in an insoluble form in basement membranes especially those underlying endothelial cells and in loose connective tissue. Fibronectin was demonstrated in cultured human endothelial cells and in the surrounding extracellular matrix by immunofluorescence microscopy by using antibody to human plasma fibronectin. Cultured human endothelial cells released fibronectin into the culture medium which was immunologically identical to the fibronectin in human plasma. Cultured human endothelial cells were labeled with [3H] leucine. The radioactive fibronectin present in the endothelial postculture medium and in urea extracts of cellular monolayers was isolated with either anti-fibronectin coupled to Protein A-Sepharose or double antibody immunoprecipitation and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When reduced, the [3H] fibronectin synthesized by cultured endothelial cells had the same mol wt (approximately 200,000) as plasma fibronectin. Unreduced, the [3H] fibronectin synthesized by endothelial cells migrated as a dimer, as did plasma fibronectin. Fibronectin accounted for approximately 15% of the protein synthesized and released by endothelial cells into the culture medium. Thus, cultured endothelial cells synthesize fibronectin, secrete it into the culture medium, and incorporate it into extracellular matrix. The results suggest that the endothelial cell is potentially a major site of synthesis of circulating plasma fibronectin. In addition, fibronectin derived from endothelial cells may be an important structural component of the subendothelium.

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Selected References

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  1. 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]
  2. Blumenstock F., Weber P., Saba T. M. Isolation and biochemical characterization of alpha-2-opsonic glycoprotein from rat serum. J Biol Chem. 1977 Oct 25;252(20):7156–7162. [PubMed] [Google Scholar]
  3. Bornstein P., Ash J. F. Cell surface-associated structural proteins in connective tissue cells. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2480–2484. doi: 10.1073/pnas.74.6.2480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Howard B. V., Macarak E. J., Gunson D., Kefalides N. A. Characterization of the collagen synthesized by endothelial cells in culture. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2361–2364. doi: 10.1073/pnas.73.7.2361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hynes R. O., Bye J. M. Density and cell cycle dependence of cell surface proteins in hamster fibroblasts. Cell. 1974 Oct;3(2):113–120. doi: 10.1016/0092-8674(74)90114-7. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hynes R. O., Destree A. Extensive disulfide bonding at the mammalian cell surface. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2855–2859. doi: 10.1073/pnas.74.7.2855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hök M., Rubin K., Oldberg A., Obrink B., Vaheri A. Cold-insoluble globulin mediates the adhesion of rat liver cells to plastic Petri dishes. Biochem Biophys Res Commun. 1977 Dec 7;79(3):726–733. doi: 10.1016/0006-291x(77)91172-x. [DOI] [PubMed] [Google Scholar]
  9. Jaffe E. A., Hoyer L. W., Nachman R. L. Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J Clin Invest. 1973 Nov;52(11):2757–2764. doi: 10.1172/JCI107471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jaffe E. A., Hoyer L. W., Nachman R. L. Synthesis of von Willebrand factor by cultured human endothelial cells. Proc Natl Acad Sci U S A. 1974 May;71(5):1906–1909. doi: 10.1073/pnas.71.5.1906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jaffe E. A., Minick C. R., Adelman B., Becker C. G., Nachman R. Synthesis of basement membrane collagen by cultured human endothelial cells. J Exp Med. 1976 Jul 1;144(1):209–225. doi: 10.1084/jem.144.1.209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jaffe E. A., Nachman R. L. Subunit structure of factor VIII antigen synthesized by cultured human endothelial cells. J Clin Invest. 1975 Sep;56(3):698–702. doi: 10.1172/JCI108140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jilek F., Hörmann H. Cold-insoluble globulin, II. Plasminolysis of cold-insoluble globulin. Hoppe Seylers Z Physiol Chem. 1977 Jan;358(1):133–136. [PubMed] [Google Scholar]
  15. 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]
  16. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  17. Klebe R. J. Isolation of a collagen-dependent cell attachment factor. Nature. 1974 Jul 19;250(463):248–251. doi: 10.1038/250248a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Linder E., Vaheri A., Ruoslahti E., Wartiovaara J. Distribution of fibroblast surface antigen in the developing chick embryo. J Exp Med. 1975 Jul 1;142(1):41–49. doi: 10.1084/jem.142.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mosesson M. W., Umfleet R. A. The cold-insoluble globulin of human plasma. I. Purification, primary characterization, and relationship to fibrinogen and other cold-insoluble fraction components. J Biol Chem. 1970 Nov 10;245(21):5728–5736. [PubMed] [Google Scholar]
  22. Mosher D. F. Action of fibrin-stabilizing factor on cold-insoluble globulin and alpha2-macroglobulin in clotting plasma. J Biol Chem. 1976 Mar 25;251(6):1639–1645. [PubMed] [Google Scholar]
  23. Mosher D. F. Cross-linking of cold-insoluble globulin by fibrin-stabilizing factor. J Biol Chem. 1975 Aug 25;250(16):6614–6621. [PubMed] [Google Scholar]
  24. Mosher D. F., Saksela O., Keski-Oja J., Vaheri A. Distribution of a major surface-associated glycoprotein, fibronectin, in cultures of adherent cells. J Supramol Struct. 1977;6(4):551–557. doi: 10.1002/jss.400060408. [DOI] [PubMed] [Google Scholar]
  25. Mosher D. F., Saksela O., Vaheri A. Synthesis and secretion of alpha-2-macroglobulin by cultured adherent lung cells. Comparison with cell strains derived from other tissues. J Clin Invest. 1977 Nov;60(5):1036–1045. doi: 10.1172/JCI108854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pearlstein E. Plasma membrane glycoprotein which mediates adhesion of fibroblasts to collagen. Nature. 1976 Aug 5;262(5568):497–500. doi: 10.1038/262497a0. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Ruoslahti E., Vaheri A., Kuusela P., Linder E. Fibroblast surface antigen: a new serum protein. Biochim Biophys Acta. 1973 Oct 18;322(2):352–358. doi: 10.1016/0005-2795(73)90310-3. [DOI] [PubMed] [Google Scholar]
  29. Schlessinger J., Barak L. S., Hammes G. G., Yamada K. M., Pastan I., Webb W. W., Elson E. L. Mobility and distribution of a cell surface glycoprotein and its interaction with other membrane components. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2909–2913. doi: 10.1073/pnas.74.7.2909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stenman S., Vaheri A. Distribution of a major connective tissue protein, fibronectin, in normal human tissues. J Exp Med. 1978 Apr 1;147(4):1054–1064. doi: 10.1084/jem.147.4.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vaheri A., Ruoslahti E. Fibroblast surface antigen produced but not retained by virus-transformed human cells. J Exp Med. 1975 Aug 1;142(2):530–535. doi: 10.1084/jem.142.2.530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weinstein M. J., Deykin D., Davie E. W. Quantitative determination of factor-VIII protein by two-stage gel electrophoresis. Br J Haematol. 1976 Jul;33(3):343–355. doi: 10.1111/j.1365-2141.1976.tb03550.x. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Yamada K. M., Yamada S. S., Pastan I. Cell surface protein partially restores morphology, adhesiveness, and contact inhibition of movement to transformed fibroblasts. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1217–1221. doi: 10.1073/pnas.73.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamada K. M., Yamada S. S., Pastan I. The major cell surface glycoprotein of chick embryo fibroblasts is an agglutinin. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3158–3162. doi: 10.1073/pnas.72.8.3158. [DOI] [PMC free article] [PubMed] [Google Scholar]

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