Skip to main content
NCBI home page
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
. 1976 Jul;73(7):2361–2364. doi: 10.1073/pnas.73.7.2361

Characterization of the collagen synthesized by endothelial cells in culture.

B V Howard, E J Macarak, D Gunson, N A Kefalides
PMCID: PMC430566  PMID: 59926

Abstract

[14C]Proline and [14C]lysine were incorporated into collagen by cultures of endothelial cells derived from calf aortae. The isomer 3-hydroxy[14C]proline accounted for 10% of the total hydroxy[14C]proline in the collagen isolated from the medium. Approximately 81% of the hydroxy[14C]lysine isolated from the medium was glycosylated, and 91% of the glycosylated hydroxy[14C]lysine was in the form of the disaccharide glucosylgalactose. Gel filtration chromatography or acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicated that the initially synthesized peptide chain of [14C]collagen had a molecular weight of about 135,000; after pepsin digestion this was converted to 115,000. The ratio of hydroxy[14C]proline to total [14C]proline x 100 in the pesin-resistant fraction was 59. When examined by immunofluorescence microscopy, the endothelial cultures stained positively with antiserum to (Type IV) collagen from basement membrane of bovine anterior lens capsule. The data indicate that cultured endothelial cells derived from calf aortae synthesize collagen that resembles that of basement membrane collagen.

Full text

PDF
2361

Images in this article

2364Image
on p.2364

Selected References

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

  1. Anker H. S. A solubilizable acrylamide gel for electrophoresis. FEBS Lett. 1970 Apr 16;7(3):293–293. doi: 10.1016/0014-5793(70)80185-5. [DOI] [PubMed] [Google Scholar]
  2. Askenasi R. S., Kefalides N. A. Simple chromatographic method for determination of 14 C-labeled lysine, hydroxylysine, and hydroxylysine glycosides. Anal Biochem. 1972 May;47(1):67–72. doi: 10.1016/0003-2697(72)90279-5. [DOI] [PubMed] [Google Scholar]
  3. Clark C. C., Tomichek E. A., Koszalka T. R., Minor R. R., Kefalides N. A. The embryonic rat parietal yolk sac. The role of the parietal endoderm in the biosynthesis of basement membrane collagen and glycoprotein in vitro. J Biol Chem. 1975 Jul 10;250(13):5259–5267. [PubMed] [Google Scholar]
  4. Cristofalo V. J., Kritchevsky D. Respiration and glycolysis in the human diploid cell strain WI-38. J Cell Physiol. 1966 Feb;67(1):125–132. doi: 10.1002/jcp.1040670114. [DOI] [PubMed] [Google Scholar]
  5. Dehm P., Prockop D. J. Biosynthesis of cartilage procollagen. Eur J Biochem. 1973 May;35(1):159–166. doi: 10.1111/j.1432-1033.1973.tb02821.x. [DOI] [PubMed] [Google Scholar]
  6. Gimbrone M. A., Jr, Cotran R. S., Folkman J. Human vascular endothelial cells in culture. Growth and DNA synthesis. J Cell Biol. 1974 Mar;60(3):673–684. doi: 10.1083/jcb.60.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Grant M. E., Harwood R., Williams I. F. The biosynthesis of basement-membrane collagen by isolated rat glomeruli. Eur J Biochem. 1975 Jun;54(2):531–540. doi: 10.1111/j.1432-1033.1975.tb04166.x. [DOI] [PubMed] [Google Scholar]
  9. Grant M. E., Kefalides N. A., Prockop D. J. The biosynthesis of basement membrane collagen in embryonic chick lens. I. Delay between the synthesis of polypeptide chains and the secretion of collagen by matrix-free cells. J Biol Chem. 1972 Jun 10;247(11):3539–3544. [PubMed] [Google Scholar]
  10. Grant M. E., Kefalides N. A., Prockop D. J. The biosynthesis of basement membrane collagen in embryonic chick lens. II. Synthesis of a precursor form by matrix-free cells and a time-dependent conversion to chains in intact lens. J Biol Chem. 1972 Jun 10;247(11):3545–3551. [PubMed] [Google Scholar]
  11. Grant M. E., Schofield J. D., Kefalides N. A., Prockop D. J. The biosynthesis of basement membrane collagen in embryonic chick lens. 3. Intracellular formation of the triple helix and the formation of aggregates through disulfide bonds. J Biol Chem. 1973 Nov 10;248(21):7432–7437. [PubMed] [Google Scholar]
  12. Howard B. V., Howard W. J., De la Llera M., Kefalides N. A. Triglyceride accumulation in cultured human fibroblasts: the effects of hypertriglyceridemic serum. Atherosclerosis. 1976 May-Jun;23(3):521–534. doi: 10.1016/0021-9150(76)90012-5. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Kefalides N. A., Cameron J. D., Tomichek E. A., Yanoff M. Biosynthesis of basement membrane collagen by rabbit corneal endothelium in vitro. J Biol Chem. 1976 Feb 10;251(3):730–733. [PubMed] [Google Scholar]
  16. Kefalides N. A., Denduchis B. Structural components of epithelial and endothelial basement membranes. Biochemistry. 1969 Nov;8(11):4613–4621. doi: 10.1021/bi00839a057. [DOI] [PubMed] [Google Scholar]
  17. Kefalides N. A. Structure and biosynthesis of basement membranes. Int Rev Connect Tissue Res. 1973;6:63–104. doi: 10.1016/b978-0-12-363706-2.50008-8. [DOI] [PubMed] [Google Scholar]
  18. Krisko I., Walker W. G. Protein and glycoprotein synthesis by isolated kidney glomeruli. Proc Soc Exp Biol Med. 1974 Jul;146(3):942–947. doi: 10.3181/00379727-146-38224. [DOI] [PubMed] [Google Scholar]
  19. 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]

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