Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 May 1;102(5):1979–1984. doi: 10.1083/jcb.102.5.1979

Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group

PMCID: PMC2114207  PMID: 3700480

Abstract

Earlier work from our laboratory demonstrated that heparin inhibited the proliferation of vascular smooth muscle cells in vivo and in vitro. Both anticoagulant and non-anticoagulant heparin species were equally effective as antiproliferative agents. Previous structure-function studies indicated that hexasaccharide and larger fragments retained antiproliferative activity, whereas tetra- and disaccharides were inactive. These experiments also suggested that both N- and O-sulfates of heparin were necessary for growth inhibitory capacity. In this paper, we have further analyzed the structural determinants of the antiproliferative activity of heparin. These experiments were done using synthetically prepared and therefore chemically defined heparin oligosaccharides. We present evidence that a pentasaccharide fragment retains antiproliferative activity, and that the 3-O-sulfate on the internal glucosamine residue is critical for growth inhibitory capacity of the pentasaccharide. We also show that heparins obtained from different manufacturers differ significantly in their ability to suppress smooth muscle cell proliferation.

Full Text

The Full Text of this article is available as a PDF (699.3 KB).

Selected References

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

  1. Burrowes C. E., Habal F. M., Movat H. Z. The inhibition of human plasma kallikrein by antithrombin III. Thromb Res. 1975 Jul;7(1):175–183. doi: 10.1016/0049-3848(75)90134-6. [DOI] [PubMed] [Google Scholar]
  2. Castellot J. J., Jr, Addonizio M. L., Rosenberg R., Karnovsky M. J. Cultured endothelial cells produce a heparinlike inhibitor of smooth muscle cell growth. J Cell Biol. 1981 Aug;90(2):372–379. doi: 10.1083/jcb.90.2.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castellot J. J., Jr, Beeler D. L., Rosenberg R. D., Karnovsky M. J. Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. J Cell Physiol. 1984 Sep;120(3):315–320. doi: 10.1002/jcp.1041200309. [DOI] [PubMed] [Google Scholar]
  4. Castellot J. J., Jr, Favreau L. V., Karnovsky M. J., Rosenberg R. D. Inhibition of vascular smooth muscle cell growth by endothelial cell-derived heparin. Possible role of a platelet endoglycosidase. J Biol Chem. 1982 Oct 10;257(19):11256–11260. [PubMed] [Google Scholar]
  5. Choay J., Petitou M., Lormeau J. C., Sinaÿ P., Casu B., Gatti G. Structure-activity relationship in heparin: a synthetic pentasaccharide with high affinity for antithrombin III and eliciting high anti-factor Xa activity. Biochem Biophys Res Commun. 1983 Oct 31;116(2):492–499. doi: 10.1016/0006-291x(83)90550-8. [DOI] [PubMed] [Google Scholar]
  6. Clowes A. W., Karnowsky M. J. Suppression by heparin of smooth muscle cell proliferation in injured arteries. Nature. 1977 Feb 17;265(5595):625–626. doi: 10.1038/265625a0. [DOI] [PubMed] [Google Scholar]
  7. Guyton J. R., Rosenberg R. D., Clowes A. W., Karnovsky M. J. Inhibition of rat arterial smooth muscle cell proliferation by heparin. In vivo studies with anticoagulant and nonanticoagulant heparin. Circ Res. 1980 May;46(5):625–634. doi: 10.1161/01.res.46.5.625. [DOI] [PubMed] [Google Scholar]
  8. Hoover R. L., Rosenberg R., Haering W., Karnovsky M. J. Inhibition of rat arterial smooth muscle cell proliferation by heparin. II. In vitro studies. Circ Res. 1980 Oct;47(4):578–583. doi: 10.1161/01.res.47.4.578. [DOI] [PubMed] [Google Scholar]
  9. Kazatchkine M. D., Fearon D. T., Metcalfe D. D., Rosenberg R. D., Austen K. F. Structural determinants of the capacity of heparin to inhibit the formation of the human amplification C3 convertase. J Clin Invest. 1981 Jan;67(1):223–228. doi: 10.1172/JCI110017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Larsen M. L., Abildgaard U., Teien A. N., Gjesdal K. Assay of plasma heparin using thrombin and the chromogenic substrate H-D-Phe-Pip-Arg-pNA (S-2238). Thromb Res. 1978 Aug;13(2):285–288. doi: 10.1016/0049-3848(78)90016-6. [DOI] [PubMed] [Google Scholar]
  11. Larson D. M., Fujiwara K., Alexander R. W., Gimbrone M. A., Jr Myosin in cultured vascular smooth muscle cells: immunofluorescence and immunochemical studies of alterations in antigenic expression. J Cell Biol. 1984 Nov;99(5):1582–1589. doi: 10.1083/jcb.99.5.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Oosta G. M., Gardner W. T., Beeler D. L., Rosenberg R. D. Multiple functional domains of the heparin molecule. Proc Natl Acad Sci U S A. 1981 Feb;78(2):829–833. doi: 10.1073/pnas.78.2.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Petitou M. Synthetic heparin fragments: new and efficient tools for the study of heparin and its interactions. Nouv Rev Fr Hematol. 1984;26(4):221–226. [PubMed] [Google Scholar]
  14. Ross R., Glomset J. A. Atherosclerosis and the arterial smooth muscle cell: Proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science. 1973 Jun 29;180(4093):1332–1339. doi: 10.1126/science.180.4093.1332. [DOI] [PubMed] [Google Scholar]
  15. Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sache E., Maillard M., Bertrand H., Maman M., Kunz M., Choay J., Fareed J., Messmore H. Studies on a highly active anticoagulant fraction of high molecular weight isolated from porcine sodium heparin. Thromb Res. 1982 Mar 15;25(6):443–458. doi: 10.1016/0049-3848(82)90086-x. [DOI] [PubMed] [Google Scholar]
  17. Venneröd A. M., Laake K. Inhibition of purified plasma kallikrein by antithrombin III and heparin. Thromb Res. 1975 Jul;7(1):223–226. doi: 10.1016/0049-3848(75)90138-3. [DOI] [PubMed] [Google Scholar]
  18. Whittemore A. D., Clowes A. W., Couch N. P., Mannick J. A. Secondary femoropopliteal reconstruction. Ann Surg. 1981 Jan;193(1):35–42. doi: 10.1097/00000658-198101000-00006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yin E. T., Wessler S., Butler J. V. Plasma heparin: a unique, practical, submicrogram-sensitive assay. J Lab Clin Med. 1973 Feb;81(2):298–310. [PubMed] [Google Scholar]

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

RESOURCES