Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1978 Jun;75(6):2621–2625. doi: 10.1073/pnas.75.6.2621

Synthesis of cold-insoluble globulin by cultured calf endothelial cells.

E J Macarak, E Kirby, T Kirk, N A Kefalides
PMCID: PMC392614  PMID: 96438

Abstract

Radiolabeled amino acids were incorporated into nondialyzable protein by cultures of endothelial cells derived from calf aorta. Antibody prepared against purified bovine plasma cold-insoluble globulin (CIG) formed a strong precipitin line upon immunodiffusion against 3H-labeled proteins from endothelial cell culture media. This precipitin line formed a line of identity with the precipitin line formed by anti-CIG and purified CIG. Double-antibody immunoprecipitation of 3H-labeled protein showed that 36.3% of the radioactivity was specifically precipitated by the anti-CIG antibody. Gel filtration and polyacrylamide gel electrophoresis of the immunoprecipitate indicated that it contained a single protein species whose molecular weight was consistent with that of CIG isolated from plasma. When cultured cells were examined by indirect immunofluorescence microscopy, the endothelial cells stained specifically with anti-CIG antibody. These data indicate that cultured endothelial cells derived from calf aorta synthesize and secrete a protein antigenically similar to CIG.

Full text

PDF
2621

Images in this article

Selected References

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

  1. Chen L. B. Alteration in cell surface LETS protein during myogenesis. Cell. 1977 Mar;10(3):393–400. doi: 10.1016/0092-8674(77)90026-5. [DOI] [PubMed] [Google Scholar]
  2. Chen L. B., Gallimore P. H., McDougall J. K. Correlation between tumor induction and the large external transformation sensitive protein on the cell surface. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3570–3574. doi: 10.1073/pnas.73.10.3570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Goldberg B., Epstein E. H., Jr, Sherr C. J. Precursors of collagen secreted by cultured human fibroblasts. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3655–3659. doi: 10.1073/pnas.69.12.3655. [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. 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]
  7. Juva K., Prockop D. J. Modified procedure for the assay of H-3-or C-14-labeled hydroxyproline. Anal Biochem. 1966 Apr;15(1):77–83. doi: 10.1016/0003-2697(66)90249-1. [DOI] [PubMed] [Google Scholar]
  8. Kuusela P., Ruoslahti E., Engvall E., Vaheri A. Immunological interspecies cross-reactions of fibroblast surface antigen (fibronectin). Immunochemistry. 1976 Aug;13(8):639–642. doi: 10.1016/0019-2791(76)90203-2. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Lewis L. J., Hoak J. C., Maca R. D., Fry G. L. Replication of human endothelial cells in culture. Science. 1973 Aug 3;181(4098):453–454. doi: 10.1126/science.181.4098.453. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. MacGregor R. R., Macarak E. J., Kefalides N. A. Comparative adherence of granulocytes to endothelial monolayers and nylon fiber. J Clin Invest. 1978 Mar;61(3):697–702. doi: 10.1172/JCI108981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Macarak E. J., Howard B. V., Kefalides N. A. Properties of calf endothelial cells in culture. Lab Invest. 1977 Jan;36(1):62–67. [PubMed] [Google Scholar]
  14. Mosesson M. W. Cold-insoluble globulin (CIg), a circulating cell surface protein. Thromb Haemost. 1977 Dec 15;38(4):742–750. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. Ruoslahti E., Vaheri A. Novel human serum protein from fibroblast plasma membrane. Nature. 1974 Apr 26;248(5451):789–791. doi: 10.1038/248789a0. [DOI] [PubMed] [Google Scholar]
  19. Schmer G., Kirby E. P., Teller D. C., Davie E. W. The isolation nd characterization of bovine factor VIII (antihemophilic factor). J Biol Chem. 1972 Apr 25;247(8):2512–2521. [PubMed] [Google Scholar]
  20. Scow R. O., Blanchette-Mackie E. J., Smith L. C. Role of capillary endothelium in the clearance of chylomicrons. A model for lipid transport from blood by lateral diffusion in cell membranes. Circ Res. 1976 Aug;39(2):149–162. doi: 10.1161/01.res.39.2.149. [DOI] [PubMed] [Google Scholar]
  21. Small R., Macarak E., Fisher A. B. Production of 5-hydroxyindoleacetic acid from serotonin by cultured endothelial cells. J Cell Physiol. 1977 Feb;90(2):225–231. doi: 10.1002/jcp.1040900208. [DOI] [PubMed] [Google Scholar]
  22. Vaheri A., Ruoslahti E., Westermark B., Ponten J. A common cell-type specific surface antigen in cultured human glial cells and fibroblasts: loss in malignant cells. J Exp Med. 1976 Jan 1;143(1):64–72. doi: 10.1084/jem.143.1.64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wasteson A., Glimelius B., Busch C., Westermark H., Heldin C. H., Norling B. Effect of a platelet endoglycosidase on cell surface associated heparan sulphate of human culturei endothelial and glial cells. Thromb Res. 1977 Sep;11(3):309–321. doi: 10.1016/0049-3848(77)90184-0. [DOI] [PubMed] [Google Scholar]
  24. Weksler B. B., Marcus A. J., Jaffe E. A. Synthesis of prostaglandin I2 (prostacyclin) by cultured human and bovine endothelial cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3922–3926. doi: 10.1073/pnas.74.9.3922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES