Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1980 Jan 15;186(1):217–225. doi: 10.1042/bj1860217

The effect of ascorbic acid on the nature and production of collagen and elastin by rat smooth-muscle cells.

Y A de Clerck, P A Jones
PMCID: PMC1161522  PMID: 7370010

Abstract

1. The effects of various concentrations of ascorbic acid on the quality and quantity of the insoluble extracellular matrices produced by two strains of cultured rat smooth-muscle cells were studied. 2. Ascorbic acid was necessary for the appearance of insoluble collagen in the extracellular matrix. 3. Secretion of soluble collagen continued in the absence of ascorbic acid, but this soluble collagen was markedly underhydroxylated. 4. The amount of insoluble collagen present in the matrix was directly related to the ascorbic acid concentration. 5. The insoluble collagen that appeared in the matrix under conditions where ascorbic acid was limiting was no more than 7% underhydroxylated. 6. In contrast, the amount of insoluble elastin produced was inversely proportional to the ascorbic acid concentration. 7. The elastin produced in the absence of ascorbic acid had the expected amino acid composition, but hydroxyproline was absent. 8. The hydroxyproline content of elastin was also directly dependent on the ascorbic acid concentration. 9. Ascorbic acid had variable effects on the quantity of glycoprotein(s) present in the matrix. 10. The appearance of insoluble collagen in the extracellular matrices produced by cultured human fibroblasts and calf endothelial cells was also completely dependent on the presence of ascorbic acid.

Full text

PDF
217

Selected References

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

  1. Anderson J. C. Glycoproteins of the connective tissue matrix. Int Rev Connect Tissue Res. 1976;7:251–322. doi: 10.1016/b978-0-12-363707-9.50012-5. [DOI] [PubMed] [Google Scholar]
  2. Barnes M. J., Constable B. J., Morton L. F., Kodicek E. Studies in vivo on the biosynthesis of collagen and elastin in ascorbic acid-deficient guinea pigs. Evidence for the formation and degradation of a partially hydroxylated collagen. Biochem J. 1970 Sep;119(3):575–585. doi: 10.1042/bj1190575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barnes M. J. Function of ascorbic acid in collagen metabolism. Ann N Y Acad Sci. 1975 Sep 30;258:264–277. doi: 10.1111/j.1749-6632.1975.tb29287.x. [DOI] [PubMed] [Google Scholar]
  4. Bates C. J., Bailey A. J., Prynne C. J., Levene C. I. The effect of ascorbic acid on the synthesis of collagen precursor secreted by 3T6 mouse fibroblasts in culture. Biochim Biophys Acta. 1972 Sep 29;278(2):372–390. doi: 10.1016/0005-2795(72)90241-3. [DOI] [PubMed] [Google Scholar]
  5. Bates C. J., Prynne C. J., Levene C. I. Ascorbate-dependent differences in the hydroxylation of proline and lysine in collagen synthesized by 3T6 fibroblasts in culture. Biochim Biophys Acta. 1972 Oct 31;278(3):610–616. doi: 10.1016/0005-2795(72)90025-6. [DOI] [PubMed] [Google Scholar]
  6. Bates C. J., Prynne C. J., Levene C. I. The synthesis of underhydroxylated collagen by 3 T6 mouse fibroblasts in culture. Biochim Biophys Acta. 1972 Apr 15;263(2):397–405. doi: 10.1016/0005-2795(72)90091-8. [DOI] [PubMed] [Google Scholar]
  7. Benya P. D., Padilla S. R., Nimni M. E. Independent regulation of collagen types by chondrocytes during the loss of differentiated function in culture. Cell. 1978 Dec;15(4):1313–1321. doi: 10.1016/0092-8674(78)90056-9. [DOI] [PubMed] [Google Scholar]
  8. Berg R. A., Prockop D. J. The thermal transition of a non-hydroxylated form of collagen. Evidence for a role for hydroxyproline in stabilizing the triple-helix of collagen. Biochem Biophys Res Commun. 1973 May 1;52(1):115–120. doi: 10.1016/0006-291x(73)90961-3. [DOI] [PubMed] [Google Scholar]
  9. Berman J., Stoner G., Dawe C., Rice J., Kingsbury E. Histochemical demonstration of collagen fibers in ascorbic-acid-fed cell cultures. In Vitro. 1978 Aug;14(8):675–685. doi: 10.1007/BF02616164. [DOI] [PubMed] [Google Scholar]
  10. Burke J. M., Ross R. Collagen synthesis by monkey arterial smooth muscle cells during proliferation and quiescence in culture. Exp Cell Res. 1977 Jul;107(2):387–395. doi: 10.1016/0014-4827(77)90360-3. [DOI] [PubMed] [Google Scholar]
  11. Cardinale G. J., Stassen F. L., Kuttan R., Udenfriend S. Activation of prolyl hydroxylase in fibroblasts by ascorbic acid. Ann N Y Acad Sci. 1975 Sep 30;258:278–287. doi: 10.1111/j.1749-6632.1975.tb29288.x. [DOI] [PubMed] [Google Scholar]
  12. Evans C. A., Peterkofsky B. Ascorbate-independent proline hydroxylation resulting from viral transformation of Balb 3T3 cells and unaffected by dibutyryl cAMP treatment. J Cell Physiol. 1976 Nov;89(3):355–367. doi: 10.1002/jcp.1040890302. [DOI] [PubMed] [Google Scholar]
  13. Faris B., Snider R., Levine A., Moscaritolo R., Salcedo L., Franzblau C. Effect of ascorbate on collagen synthesis by lung embryonic fibroblasts. In Vitro. 1978 Dec;14(12):1022–1027. doi: 10.1007/BF02616217. [DOI] [PubMed] [Google Scholar]
  14. Fessler J. H., Fessler L. I. Biosynthesis of procollagen. Annu Rev Biochem. 1978;47:129–162. doi: 10.1146/annurev.bi.47.070178.001021. [DOI] [PubMed] [Google Scholar]
  15. GOLDBERG B., GREEN H., TODARO G. J. COLLAGEN FORMATION IN VITRO BY ESTABLISHED MAMMALIAN CELL LINES. Exp Cell Res. 1963 Aug;31:444–447. doi: 10.1016/0014-4827(63)90025-9. [DOI] [PubMed] [Google Scholar]
  16. GREEN H., GOLDBERG B. COLLAGEN SYNTHESIS BY HUMAN FIBROBLAST STRAINS. Proc Soc Exp Biol Med. 1964 Oct;117:258–261. doi: 10.3181/00379727-117-29551. [DOI] [PubMed] [Google Scholar]
  17. Gallop P. M., Blumenfeld O. O., Seifter S. Structure and metabolism of connective 801 tissue proteins. Annu Rev Biochem. 1972;41:617–672. doi: 10.1146/annurev.bi.41.070172.003153. [DOI] [PubMed] [Google Scholar]
  18. Gay S., Martin G. R., Muller P. K., Timpl R., Kuhn K. Simultaneous synthesis of types I and III collagen by fibroblasts in culture. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4037–4040. doi: 10.1073/pnas.73.11.4037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gimbrone M. A., Jr, Cotran R. S. Human vascular smooth muscle in culture. Growth and ultrastructure. Lab Invest. 1975 Jul;33(1):16–27. [PubMed] [Google Scholar]
  20. Grant M. E., Prockop D. J. The biosynthesis of collagen. 1. N Engl J Med. 1972 Jan 27;286(4):194–199. doi: 10.1056/NEJM197201272860406. [DOI] [PubMed] [Google Scholar]
  21. Grant M. E., Prockop D. J. The biosynthesis of collagen. 2. N Engl J Med. 1972 Feb 3;286(5):242–249. doi: 10.1056/NEJM197202032860505. [DOI] [PubMed] [Google Scholar]
  22. Grant M. E., Prockop D. J. The biosynthesis of collagen. 3. N Engl J Med. 1972 Feb 10;286(6):291–300. doi: 10.1056/NEJM197202102860604. [DOI] [PubMed] [Google Scholar]
  23. Gribble T. J., Comstock J. P., Udenfriend S. Collagen chain formation and peptidyl proline hydroxylation in monolayer tissue cultures of L-929 fibroblasts. Arch Biochem Biophys. 1969 Jan;129(1):308–316. doi: 10.1016/0003-9861(69)90180-5. [DOI] [PubMed] [Google Scholar]
  24. Heinegård D., Hascall V. C. Aggregation of cartilage proteoglycans. 3. Characteristics of the proteins isolated from trypsin digests of aggregates. J Biol Chem. 1974 Jul 10;249(13):4250–4256. [PubMed] [Google Scholar]
  25. 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]
  26. Jones P. A. Construction of an artificial blood vessel wall from cultured endothelial and smooth muscle cells. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1882–1886. doi: 10.1073/pnas.76.4.1882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jones P. A., Scott-Burden T. Activated macrophages digest the extracellular matrix proteins produced by cultured cells. Biochem Biophys Res Commun. 1979 Jan 15;86(1):71–77. doi: 10.1016/0006-291x(79)90383-8. [DOI] [PubMed] [Google Scholar]
  28. Jones P. A., Scott-Burden T., Gevers W. Glycoprotein, elastin, and collagen secretion by rat smooth muscle cells. Proc Natl Acad Sci U S A. 1979 Jan;76(1):353–357. doi: 10.1073/pnas.76.1.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Kao W. W., Berg R. A., Prockop D. J. Kinetics for the secretion of procollagen by freshly isolated tendon cells. J Biol Chem. 1977 Dec 10;252(23):8391–8397. [PubMed] [Google Scholar]
  31. Levene C. I., Bates C. J. Ascorbic acid and collagen synthesis in cultured fibroblasts. Ann N Y Acad Sci. 1975 Sep 30;258:288–306. doi: 10.1111/j.1749-6632.1975.tb29289.x. [DOI] [PubMed] [Google Scholar]
  32. Mayne R., Vail M. S., Miller E. J. Characterization of the collagen chains synthesized by cultured smooth muscle cells derived from rhesus monkey thoracic aorta. Biochemistry. 1978 Feb 7;17(3):446–452. doi: 10.1021/bi00596a011. [DOI] [PubMed] [Google Scholar]
  33. Peterkofsky B., Prather W. B. Increased collagen synthesis in Kirsten sarcoma virus-transformed BALB 3T3 cells grown in the presence of dibutyryl cyclic AMP. Cell. 1974 Nov;3(3):291–299. doi: 10.1016/0092-8674(74)90144-5. [DOI] [PubMed] [Google Scholar]
  34. Rucker R. B., Tinker D. Structure and metabolism of arterial elastin. Int Rev Exp Pathol. 1977;17:1–47. [PubMed] [Google Scholar]
  35. Schwarz R. I., Bissell M. J. Dependence of the differentiated state on the cellular environment: modulation of collagen synthesis in tendon cells. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4453–4457. doi: 10.1073/pnas.74.10.4453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sear C. H., Grant M. E., Jackson D. S. Biosynthesis and release of glycoproteins by human skin fibroblasts in culture. Biochem J. 1977 Oct 15;168(1):91–103. doi: 10.1042/bj1680091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sykes B. C., Partridge S. M. Isolation of a soluble elastin from lathyritic chicks. Biochem J. 1972 Dec;130(4):1171–1172. doi: 10.1042/bj1301171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tanzer M. L. Cross-linking of collagen. Science. 1973 May 11;180(4086):561–566. doi: 10.1126/science.180.4086.561. [DOI] [PubMed] [Google Scholar]
  39. Uitto J., Hoffmann H-P, Prockop D. J. Synthesis of elastin and procallagen by cells from embryonic aorta. Differences in the role of hydroxyproline and the effects of proline analogs on the secretion of the two proteins. Arch Biochem Biophys. 1976 Mar;173(1):187–200. doi: 10.1016/0003-9861(76)90249-6. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES