Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1990 Apr 1;267(1):185–189. doi: 10.1042/bj2670185

Release of hyaluronate from eukaryotic cells.

P Prehm 1
PMCID: PMC1131262  PMID: 2158307

Abstract

The mechanism of hyaluronate shedding from eukaryotic cell lines was analysed. All cell lines shed identical sizes of hyaluronate as were retained on the surface. They differed in the amount of hyaluronate synthesized and in the proportions of hyaluronate which were released and retained. A method was developed which could discriminate between shedding due to intramolecular degradation and that due to dissociation as intact macromolecules. This method was applied to B6 and SV3T3 cells in order to study the mechanism of hyaluronate release in more detail. The cells were pulse-labelled to form hyaluronate chains with labelled and unlabelled segments, and the sizes of labelled hyaluronate released into the medium during the pulse extension period were determined by gel filtration. B6 cells released identical sizes of hyaluronate at all labelled segment lengths, indicating that no intramolecular degradation occurred. When chain elongation was blocked by periodate-oxidized UDP-glucuronic acid, hyaluronate release was simultaneously inhibited. These results indicated that B6 cells dissociated hyaluronate as an intact macromolecule. In contrast, SV3T3 cells released hyaluronate of varying molecular mass distributions during extension of the labelled segment, suggesting partial degradation. Exogenous hyaluronate added to SV3T3 cultures was also degraded. This degradation could be prevented by the presence of radical scavengers such as superoxide dismutase and tocopherol. Degradation of endogenous hyaluronate could be inhibited by salicylate. These results led to the conclusion that SV3T3 cells released hyaluronate not only by dissociation, but also by radical-induced degradation.

Full text

PDF
185

Selected References

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

  1. Bernanke D. H., Orkin R. W. Hyaluronidase activity in embryonic chick heart muscle and cushion tissue and cells. Dev Biol. 1984 Dec;106(2):351–359. doi: 10.1016/0012-1606(84)90233-1. [DOI] [PubMed] [Google Scholar]
  2. Brecht M., Mayer U., Schlosser E., Prehm P. Increased hyaluronate synthesis is required for fibroblast detachment and mitosis. Biochem J. 1986 Oct 15;239(2):445–450. doi: 10.1042/bj2390445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clarris B. J., Fraser J. R. On the pericellular zone of some mammalian cells in vitro. Exp Cell Res. 1968 Jan;49(1):181–193. doi: 10.1016/0014-4827(68)90530-2. [DOI] [PubMed] [Google Scholar]
  4. Goldberg R. L., Toole B. P. Hyaluronate inhibition of cell proliferation. Arthritis Rheum. 1987 Jul;30(7):769–778. doi: 10.1002/art.1780300707. [DOI] [PubMed] [Google Scholar]
  5. Klein U., von Figura K. Characterization of dermatan sulfate in mucopolysaccharidosis VI. Evidence for the absence of hyaluronidase-like enzymes in human skin fibroblasts. Biochim Biophys Acta. 1980 Jun 5;630(1):10–14. doi: 10.1016/0304-4165(80)90131-2. [DOI] [PubMed] [Google Scholar]
  6. McCord J. M. Free radicals and inflammation: protection of synovial fluid by superoxide dismutase. Science. 1974 Aug 9;185(4150):529–531. doi: 10.1126/science.185.4150.529. [DOI] [PubMed] [Google Scholar]
  7. Mikuni-Takagaki Y., Toole B. P. Shedding of hyaluronate from the cell surface of Rous sarcoma virus-transformed chondrocytes. J Biol Chem. 1979 Sep 10;254(17):8409–8415. [PubMed] [Google Scholar]
  8. Oberley L. W., Buettner G. R. Role of superoxide dismutase in cancer: a review. Cancer Res. 1979 Apr;39(4):1141–1149. [PubMed] [Google Scholar]
  9. Orkin R. W., Underhill C. B., Toole B. P. Hyaluronate degradation in 3T3 and simian virus-transformed 3T3 cells. J Biol Chem. 1982 May 25;257(10):5821–5826. [PubMed] [Google Scholar]
  10. Philipson L. H., Westley J., Schwartz N. B. Effect of hyaluronidase treatment of intact cells on hyaluronate synthetase activity. Biochemistry. 1985 Dec 31;24(27):7899–7906. doi: 10.1021/bi00348a008. [DOI] [PubMed] [Google Scholar]
  11. Prehm P. Hyaluronate is synthesized at plasma membranes. Biochem J. 1984 Jun 1;220(2):597–600. doi: 10.1042/bj2200597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Prehm P. Inhibition of hyaluronate synthesis. Biochem J. 1985 Feb 1;225(3):699–705. doi: 10.1042/bj2250699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Prehm P. Synthesis of hyaluronate in differentiated teratocarcinoma cells. Characterization of the synthase. Biochem J. 1983 Apr 1;211(1):181–189. doi: 10.1042/bj2110181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Prehm P. Synthesis of hyaluronate in differentiated teratocarcinoma cells. Mechanism of chain growth. Biochem J. 1983 Apr 1;211(1):191–198. doi: 10.1042/bj2110191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Turley E. A., Moore D., Hayden L. J. Characterization of hyaluronate binding proteins isolated from 3T3 and murine sarcoma virus transformed 3T3 cells. Biochemistry. 1987 Jun 2;26(11):2997–3005. doi: 10.1021/bi00385a007. [DOI] [PubMed] [Google Scholar]
  16. Underhill C. B., Green S. J., Comoglio P. M., Tarone G. The hyaluronate receptor is identical to a glycoprotein of Mr 85,000 (gp85) as shown by a monoclonal antibody that interferes with binding activity. J Biol Chem. 1987 Sep 25;262(27):13142–13146. [PubMed] [Google Scholar]
  17. Underhill C. B., Toole B. P. Transformation-dependent loss of the hyaluronate-containing coats of cultured cells. J Cell Physiol. 1982 Feb;110(2):123–128. doi: 10.1002/jcp.1041100204. [DOI] [PubMed] [Google Scholar]
  18. Underhill C. B., Toole B. P. Transformation-dependent loss of the hyaluronate-containing coats of cultured cells. J Cell Physiol. 1982 Feb;110(2):123–128. doi: 10.1002/jcp.1041100204. [DOI] [PubMed] [Google Scholar]
  19. West D. C., Kumar S. The effect of hyaluronate and its oligosaccharides on endothelial cell proliferation and monolayer integrity. Exp Cell Res. 1989 Jul;183(1):179–196. doi: 10.1016/0014-4827(89)90428-x. [DOI] [PubMed] [Google Scholar]
  20. Wong S. F., Halliwell B., Richmond R., Skowroneck W. R. The role of superoxide and hydroxyl radicals in the degradation of hyaluronic acid induced by metal ions and by ascorbic acid. J Inorg Biochem. 1981 Apr;14(2):127–134. doi: 10.1016/s0162-0134(00)80033-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES