Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1978 Sep 1;78(3):701–715. doi: 10.1083/jcb.78.3.701

Amniotic fluid fibronectin. Characterization and synthesis by cells in culture

PMCID: PMC2110185  PMID: 701356

Abstract

A glycoprotein immunologically related to plasma cold-insoluble globulin (CIG) and fetal skin fibroblast fibronectin has been purified from second-trimester human amniotic fluid. This protein (amniotic fluid fibronectin) migrated more slowly than CIG on sodium dodecyl sulfate gel electrophoresis and showed greater polydispersity which could result, at least in part, from heterogeneity in glycosylation. Cloned human amniotic fluid epithelioid and fibroblastic cells synthesized and secreted a protein with similar properties into the culture medium. Fibronectin was shown to be associated with the pericellular and extracellular matrix of cultured amniotic fluid cells by immunofluorescence, lactoperoxidase-catalyzed iodination, and labeling with ferritin-conjugated antibodies. The kinetics of secretion of the protein were consistent with its role as a matrix protein. We anticipate that amniotic fluid fibronectin will prove to be the same protein which elsewhere in the body is incorporated into connective tissues and basement membranes. Amniotic fluid could, therefore, serve as a convenient source of in vivo synthesized fibronectin for biological and structural studies.

Full Text

The Full Text of this article is available as a PDF (2.6 MB).

Selected References

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

  1. Baum B. J., McDonald J. A., Crystal R. G. Metabolic fate of the major cell surface protein of normal human fibroblasts. Biochem Biophys Res Commun. 1977 Nov 7;79(1):8–15. doi: 10.1016/0006-291x(77)90053-5. [DOI] [PubMed] [Google Scholar]
  2. 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]
  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 A. B., Mosesson M. W., Solish G. I. Identification of the cold-insoluble globulin of plasma in amniotic fluid. Am J Obstet Gynecol. 1976 Aug 1;125(7):958–961. doi: 10.1016/0002-9378(76)90495-6. [DOI] [PubMed] [Google Scholar]
  5. Chen L. B., Maitland N., Gallimore P. H., McDougall J. K. Detection of the large external transformation-sensitive protein on some epithelial cells. Exp Cell Res. 1977 Apr;106(1):39–46. doi: 10.1016/0014-4827(77)90238-5. [DOI] [PubMed] [Google Scholar]
  6. Davidson J. M., McEneany L. S., Bornstein P. Intermediates in the conversion of procollagen to collagen. Evidence for stepwise limited proteolysis of the COOH-terminal peptide extensions. Eur J Biochem. 1977 Dec 1;81(2):349–355. doi: 10.1111/j.1432-1033.1977.tb11958.x. [DOI] [PubMed] [Google Scholar]
  7. Furcht L. T., Mosher D. F., Wendelschafer-Crabb G. Immunocytochemical localization of fibronectin (LETS proteins) on the surface of L6 myoblasts: light and electron microscopic studies. Cell. 1978 Feb;13(2):263–271. doi: 10.1016/0092-8674(78)90195-2. [DOI] [PubMed] [Google Scholar]
  8. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  9. Hoehn H., Bryant E. M., Fantel A. G., Martin G. M. Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. III. The fetal urine as a potential source of clonable cells. Humangenetik. 1975 Oct 7;29(4):285–290. doi: 10.1007/BF00394190. [DOI] [PubMed] [Google Scholar]
  10. Hoehn H., Bryant E. M., Karp L. E., Martin G. M. Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. I. Clonal morphology and growth potential. Pediatr Res. 1974 Aug;8(8):746–754. doi: 10.1203/00006450-197408000-00003. [DOI] [PubMed] [Google Scholar]
  11. Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Kishida Y., Olsen B. R., Berg R. A., Prockop D. J. Two improved methods for preparing ferritin-protein conjugates for electron microscopy. J Cell Biol. 1975 Feb;64(2):331–339. doi: 10.1083/jcb.64.2.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kruse N. J., Bornstein P. The metabolic requirements for transcellular movement and secretion of collagen. J Biol Chem. 1975 Jul 10;250(13):4841–4847. [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  19. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  20. Megaw J. M., Priest J. H., Priest R. E., Johnson L. D. Differentiation in human amniotic fluid cell cultures: II: Secretion of an epithelial basement membrane glycoprotein. J Med Genet. 1977 Jun;14(3):163–167. doi: 10.1136/jmg.14.3.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mosher D. F. Labeling of a major fibroblast surface protein (fibronectin) catalyzed by blood coagulation factor XIIa. Biochim Biophys Acta. 1977 Mar 28;491(1):205–210. doi: 10.1016/0005-2795(77)90056-3. [DOI] [PubMed] [Google Scholar]
  22. Olden K., Yamada K. M. Mechanism of the decrease in the major cell surface protein of chick embryo fibroblasts after transformation. Cell. 1977 Aug;11(4):957–969. doi: 10.1016/0092-8674(77)90307-5. [DOI] [PubMed] [Google Scholar]
  23. Priest R. E., Priest J. H., Moinuddin J. F., Keyser A. J. Differentiation in human amniotic fluid cell cultures: I: Collagen production. J Med Genet. 1977 Jun;14(3):157–162. doi: 10.1136/jmg.14.3.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Wartiovaara J., Linder E., Ruoslahti E., Vaheri A. Distribution of fibroblast surface antigen: association with fibrillar structures of normal cells and loss upon viral transformation. J Exp Med. 1974 Dec 1;140(6):1522–1533. doi: 10.1084/jem.140.6.1522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yamada K. M., Schlesinger D. H., Kennedy D. W., Pastan I. Characterization of a major fibroblast cell surface glycoprotein. Biochemistry. 1977 Dec 13;16(25):5552–5559. doi: 10.1021/bi00644a025. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Yamada K. M., Weston J. A. The synthesis, turnover, and artificial restoration of a major cell surface glycoprotein. Cell. 1975 May;5(1):75–81. doi: 10.1016/0092-8674(75)90094-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES