Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 Aug 15;350(Pt 1):291–298.

Thrombospondin 1 does not activate transforming growth factor beta1 in a chemically defined system or in smooth-muscle-cell cultures.

D J Grainger 1, E K Frow 1
PMCID: PMC1221254  PMID: 10926856

Abstract

The cytokine transforming growth factor beta1 (TGF-beta1) is secreted in a latent form that has no known biological activity. The conversion of latent TGF-beta1 into its biologically active 25 kDa form is thought to be an important step in the regulation of TGF-beta activity both in cell culture and in vivo. Thrombospondin (TSP)-1, a 360 kDa platelet alpha-granule and extracellular matrix protein, has been shown to participate in TGF-beta1 activation. We have used a chemically defined system to examine the mechanism of TSP-1-mediated TGF-beta1 activation. However, the addition of two different preparations of TSP-1 to recombinant small latent TGF-beta1 in the test tube resulted in only a very small increase in the proportion of the TGF-beta1 able to bind to the TGF-beta type II receptor: from 0.1% to a maximum of 0.4%. This small effect was not specific for TSP-1: matrix metalloproteinase 2, tissue inhibitor of matrix metalloproteinase 2 and active plasminogen activator inhibitor 1, but not transglutaminase, human serum albumin or immunoglobulin, had quantitatively similar effects on latent TGF-beta1. Furthermore, no change in the activity associated with small latent TGF-beta1 was noted in either mink lung epithelial cell or rat aortic smooth-muscle cell culture systems in the presence of TSP-1 (or TSP-1-derived peptides). We conclude that TSP-1, either alone or in the presence of cultured smooth-muscle cells (a cell type known to activate latent TGF-beta in vitro and in vivo) is unable to activate latent TGF-beta1. Any TSP-mediated activation of TGF-beta1 must depend on additional factor(s) not present in our systems.

Full Text

The Full Text of this article is available as a PDF (280.4 KB).

Selected References

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

  1. Albo D., Berger D. H., Wang T. N., Hu X., Rothman V., Tuszynski G. P. Thrombospondin-1 and transforming growth factor-beta l promote breast tumor cell invasion through up-regulation of the plasminogen/plasmin system. Surgery. 1997 Aug;122(2):493–500. doi: 10.1016/s0039-6060(97)90043-x. [DOI] [PubMed] [Google Scholar]
  2. Bailly S., Brand C., Chambaz E. M., Feige J. J. Analysis of small latent transforming growth factor-beta complex formation and dissociation by surface plasmon resonance. Absence of direct interaction with thrombospondins. J Biol Chem. 1997 Jun 27;272(26):16329–16334. doi: 10.1074/jbc.272.26.16329. [DOI] [PubMed] [Google Scholar]
  3. Border W. A., Ruoslahti E. Transforming growth factor-beta in disease: the dark side of tissue repair. J Clin Invest. 1992 Jul;90(1):1–7. doi: 10.1172/JCI115821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark D. A., Coker R. Transforming growth factor-beta (TGF-beta). Int J Biochem Cell Biol. 1998 Mar;30(3):293–298. doi: 10.1016/s1357-2725(97)00128-3. [DOI] [PubMed] [Google Scholar]
  5. Clezardin P., McGregor J. L., Manach M., Robert F., Dechavanne M., Clemetson K. J. Isolation of thrombospondin released from thrombin-stimulated human platelets by fast protein liquid chromatography on an anion-exchange Mono-Q column. J Chromatogr. 1984 Jul 27;296:249–256. doi: 10.1016/s0021-9673(01)96418-0. [DOI] [PubMed] [Google Scholar]
  6. Crawford S. E., Stellmach V., Murphy-Ullrich J. E., Ribeiro S. M., Lawler J., Hynes R. O., Boivin G. P., Bouck N. Thrombospondin-1 is a major activator of TGF-beta1 in vivo. Cell. 1998 Jun 26;93(7):1159–1170. doi: 10.1016/s0092-8674(00)81460-9. [DOI] [PubMed] [Google Scholar]
  7. Danielpour D., Dart L. L., Flanders K. C., Roberts A. B., Sporn M. B. Immunodetection and quantitation of the two forms of transforming growth factor-beta (TGF-beta 1 and TGF-beta 2) secreted by cells in culture. J Cell Physiol. 1989 Jan;138(1):79–86. doi: 10.1002/jcp.1041380112. [DOI] [PubMed] [Google Scholar]
  8. Dubois C. M., Laprise M. H., Blanchette F., Gentry L. E., Leduc R. Processing of transforming growth factor beta 1 precursor by human furin convertase. J Biol Chem. 1995 May 5;270(18):10618–10624. doi: 10.1074/jbc.270.18.10618. [DOI] [PubMed] [Google Scholar]
  9. Grainger D. J., Hesketh T. R., Metcalfe J. C., Weissberg P. L. A large accumulation of non-muscle myosin occurs at first entry into M phase in rat vascular smooth-muscle cells. Biochem J. 1991 Jul 1;277(Pt 1):145–151. doi: 10.1042/bj2770145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grainger D. J., Kemp P. R., Liu A. C., Lawn R. M., Metcalfe J. C. Activation of transforming growth factor-beta is inhibited in transgenic apolipoprotein(a) mice. Nature. 1994 Aug 11;370(6489):460–462. doi: 10.1038/370460a0. [DOI] [PubMed] [Google Scholar]
  11. Grainger D. J., Kemp P. R., Witchell C. M., Weissberg P. L., Metcalfe J. C. Transforming growth factor beta decreases the rate of proliferation of rat vascular smooth muscle cells by extending the G2 phase of the cell cycle and delays the rise in cyclic AMP before entry into M phase. Biochem J. 1994 Apr 1;299(Pt 1):227–235. doi: 10.1042/bj2990227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grainger D. J., Kirschenlohr H. L., Metcalfe J. C., Weissberg P. L., Wade D. P., Lawn R. M. Proliferation of human smooth muscle cells promoted by lipoprotein(a). Science. 1993 Jun 11;260(5114):1655–1658. doi: 10.1126/science.8503012. [DOI] [PubMed] [Google Scholar]
  13. Grainger D. J., Metcalfe J. C. A pivotal role for TGF-beta in atherogenesis? Biol Rev Camb Philos Soc. 1995 Nov;70(4):571–596. doi: 10.1111/j.1469-185x.1995.tb01652.x. [DOI] [PubMed] [Google Scholar]
  14. Grainger D. J., Wakefield L., Bethell H. W., Farndale R. W., Metcalfe J. C. Release and activation of platelet latent TGF-beta in blood clots during dissolution with plasmin. Nat Med. 1995 Sep;1(9):932–937. doi: 10.1038/nm0995-932. [DOI] [PubMed] [Google Scholar]
  15. Kanzaki T., Olofsson A., Morén A., Wernstedt C., Hellman U., Miyazono K., Claesson-Welsh L., Heldin C. H. TGF-beta 1 binding protein: a component of the large latent complex of TGF-beta 1 with multiple repeat sequences. Cell. 1990 Jun 15;61(6):1051–1061. doi: 10.1016/0092-8674(90)90069-q. [DOI] [PubMed] [Google Scholar]
  16. Kirschenlohr H. L., Metcalfe J. C., Weissberg P. L., Grainger D. J. Adult human aortic smooth muscle cells in culture produce active TGF-beta. Am J Physiol. 1993 Aug;265(2 Pt 1):C571–C576. doi: 10.1152/ajpcell.1993.265.2.C571. [DOI] [PubMed] [Google Scholar]
  17. Krutzsch H. C., Choe B. J., Sipes J. M., Guo N. h., Roberts D. D. Identification of an alpha(3)beta(1) integrin recognition sequence in thrombospondin-1. J Biol Chem. 1999 Aug 20;274(34):24080–24086. doi: 10.1074/jbc.274.34.24080. [DOI] [PubMed] [Google Scholar]
  18. Kulkarni A. B., Huh C. G., Becker D., Geiser A., Lyght M., Flanders K. C., Roberts A. B., Sporn M. B., Ward J. M., Karlsson S. Transforming growth factor beta 1 null mutation in mice causes excessive inflammatory response and early death. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):770–774. doi: 10.1073/pnas.90.2.770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lyons R. M., Gentry L. E., Purchio A. F., Moses H. L. Mechanism of activation of latent recombinant transforming growth factor beta 1 by plasmin. J Cell Biol. 1990 Apr;110(4):1361–1367. doi: 10.1083/jcb.110.4.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Munger J. S., Harpel J. G., Gleizes P. E., Mazzieri R., Nunes I., Rifkin D. B. Latent transforming growth factor-beta: structural features and mechanisms of activation. Kidney Int. 1997 May;51(5):1376–1382. doi: 10.1038/ki.1997.188. [DOI] [PubMed] [Google Scholar]
  21. Munger J. S., Huang X., Kawakatsu H., Griffiths M. J., Dalton S. L., Wu J., Pittet J. F., Kaminski N., Garat C., Matthay M. A. The integrin alpha v beta 6 binds and activates latent TGF beta 1: a mechanism for regulating pulmonary inflammation and fibrosis. Cell. 1999 Feb 5;96(3):319–328. doi: 10.1016/s0092-8674(00)80545-0. [DOI] [PubMed] [Google Scholar]
  22. Murphy-Ullrich J. E., Schultz-Cherry S., Hök M. Transforming growth factor-beta complexes with thrombospondin. Mol Biol Cell. 1992 Feb;3(2):181–188. doi: 10.1091/mbc.3.2.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pepper M. S. Transforming growth factor-beta: vasculogenesis, angiogenesis, and vessel wall integrity. Cytokine Growth Factor Rev. 1997 Mar;8(1):21–43. doi: 10.1016/s1359-6101(96)00048-2. [DOI] [PubMed] [Google Scholar]
  24. Ploplis V. A., Carmeliet P., Vazirzadeh S., Van Vlaenderen I., Moons L., Plow E. F., Collen D. Effects of disruption of the plasminogen gene on thrombosis, growth, and health in mice. Circulation. 1995 Nov 1;92(9):2585–2593. doi: 10.1161/01.cir.92.9.2585. [DOI] [PubMed] [Google Scholar]
  25. Prud'homme G. J., Piccirillo C. A. The inhibitory effects of transforming growth factor-beta-1 (TGF-beta1) in autoimmune diseases. J Autoimmun. 2000 Feb;14(1):23–42. doi: 10.1006/jaut.1999.0339. [DOI] [PubMed] [Google Scholar]
  26. Reiss M. Transforming growth factor-beta and cancer: a love-hate relationship? Oncol Res. 1997;9(9):447–457. [PubMed] [Google Scholar]
  27. Ribeiro S. M., Poczatek M., Schultz-Cherry S., Villain M., Murphy-Ullrich J. E. The activation sequence of thrombospondin-1 interacts with the latency-associated peptide to regulate activation of latent transforming growth factor-beta. J Biol Chem. 1999 May 7;274(19):13586–13593. doi: 10.1074/jbc.274.19.13586. [DOI] [PubMed] [Google Scholar]
  28. Roberts A. B., Anzano M. A., Wakefield L. M., Roche N. S., Stern D. F., Sporn M. B. Type beta transforming growth factor: a bifunctional regulator of cellular growth. Proc Natl Acad Sci U S A. 1985 Jan;82(1):119–123. doi: 10.1073/pnas.82.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schultz-Cherry S., Chen H., Mosher D. F., Misenheimer T. M., Krutzsch H. C., Roberts D. D., Murphy-Ullrich J. E. Regulation of transforming growth factor-beta activation by discrete sequences of thrombospondin 1. J Biol Chem. 1995 Mar 31;270(13):7304–7310. doi: 10.1074/jbc.270.13.7304. [DOI] [PubMed] [Google Scholar]
  30. Schultz-Cherry S., Lawler J., Murphy-Ullrich J. E. The type 1 repeats of thrombospondin 1 activate latent transforming growth factor-beta. J Biol Chem. 1994 Oct 28;269(43):26783–26788. [PubMed] [Google Scholar]
  31. Schultz-Cherry S., Murphy-Ullrich J. E. Thrombospondin causes activation of latent transforming growth factor-beta secreted by endothelial cells by a novel mechanism. J Cell Biol. 1993 Aug;122(4):923–932. doi: 10.1083/jcb.122.4.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schultz-Cherry S., Ribeiro S., Gentry L., Murphy-Ullrich J. E. Thrombospondin binds and activates the small and large forms of latent transforming growth factor-beta in a chemically defined system. J Biol Chem. 1994 Oct 28;269(43):26775–26782. [PubMed] [Google Scholar]
  33. Shull M. M., Ormsby I., Kier A. B., Pawlowski S., Diebold R. J., Yin M., Allen R., Sidman C., Proetzel G., Calvin D. Targeted disruption of the mouse transforming growth factor-beta 1 gene results in multifocal inflammatory disease. Nature. 1992 Oct 22;359(6397):693–699. doi: 10.1038/359693a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Slane J. M., Mosher D. F., Lai C. S. Conformational change in thrombospondin induced by removal of bound Ca2+. A spin label approach. FEBS Lett. 1988 Mar 14;229(2):363–366. doi: 10.1016/0014-5793(88)81157-8. [DOI] [PubMed] [Google Scholar]
  35. Tsang M. L., Zhou L., Zheng B. L., Wenker J., Fransen G., Humphrey J., Smith J. M., O'Connor-McCourt M., Lucas R., Weatherbee J. A. Characterization of recombinant soluble human transforming growth factor-beta receptor type II (rhTGF-beta sRII). Cytokine. 1995 Jul;7(5):389–397. doi: 10.1006/cyto.1995.0054. [DOI] [PubMed] [Google Scholar]

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

RESOURCES