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. 1991 Jul 1;277(Pt 1):165–173. doi: 10.1042/bj2770165

Activation of human platelet-derived latent transforming growth factor-beta 1 by human glioblastoma cells. Comparison with proteolytic and glycosidic enzymes.

D Huber 1, A Fontana 1, S Bodmer 1
PMCID: PMC1151206  PMID: 1830205

Abstract

Transforming growth factor-beta (TGF-beta), a regulator of cell growth and differentiation, is secreted by most cultured cells in latent form (L-TGF-beta). Activation of L-TGF-beta can be achieved by various physico-chemical treatments, including acidification, alkalinization, heating and chaotropic agents. Proposed physiological activators include proteinases and glycosidases, which, however, only lead to limited activation (15-20% of the total TGF-beta activity after acidic activation). In the present study L-TGF-beta 1 partially purified from human platelets was not activated by treatment with neuraminidase or the proteinases plasmin, endoproteinase Arg-C, elastase and chymotrypsin. The mechanism of activation of L-TGF-beta was further assessed by using the human glioblastoma cell line 308, which releases biologically active TGF-beta 2. Factor(s) secreted by 308 glioblastoma cells were found to be able to activate partially purified L-TGF-beta 1 from human platelets. Our finding may prove to constitute a physiologically relevant mechanism for the activation of latent forms of TGF-beta in vivo.

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Selected References

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  1. Antonelli-Orlidge A., Saunders K. B., Smith S. R., D'Amore P. A. An activated form of transforming growth factor beta is produced by cocultures of endothelial cells and pericytes. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4544–4548. doi: 10.1073/pnas.86.12.4544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BUNDY H. F. A new spectrophotometric method for the determination of chymotrypsin activity. Anal Biochem. 1962 May;3:431–435. doi: 10.1016/0003-2697(62)90072-6. [DOI] [PubMed] [Google Scholar]
  3. Bodmer S., Strommer K., Frei K., Siepl C., de Tribolet N., Heid I., Fontana A. Immunosuppression and transforming growth factor-beta in glioblastoma. Preferential production of transforming growth factor-beta 2. J Immunol. 1989 Nov 15;143(10):3222–3229. [PubMed] [Google Scholar]
  4. Brown P. D., Wakefield L. M., Levinson A. D., Sporn M. B. Physicochemical activation of recombinant latent transforming growth factor-beta's 1, 2, and 3. Growth Factors. 1990;3(1):35–43. doi: 10.3109/08977199009037500. [DOI] [PubMed] [Google Scholar]
  5. Brunner A. M., Gentry L. E., Cooper J. A., Purchio A. F. Recombinant type 1 transforming growth factor beta precursor produced in Chinese hamster ovary cells is glycosylated and phosphorylated. Mol Cell Biol. 1988 May;8(5):2229–2232. doi: 10.1128/mcb.8.5.2229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brunner A. M., Marquardt H., Malacko A. R., Lioubin M. N., Purchio A. F. Site-directed mutagenesis of cysteine residues in the pro region of the transforming growth factor beta 1 precursor. Expression and characterization of mutant proteins. J Biol Chem. 1989 Aug 15;264(23):13660–13664. [PubMed] [Google Scholar]
  7. Caltabiano M. M., Tsang M. L., Weatherbee J. A., Lucas R., Sathe G., Sutton J., Johnson G. D., Bergsma D. J. Transient production and secretion of human transforming growth factor TGF-beta 2. Gene. 1989 Dec 28;85(2):479–488. doi: 10.1016/0378-1119(89)90442-3. [DOI] [PubMed] [Google Scholar]
  8. Cheifetz S., Weatherbee J. A., Tsang M. L., Anderson J. K., Mole J. E., Lucas R., Massagué J. The transforming growth factor-beta system, a complex pattern of cross-reactive ligands and receptors. Cell. 1987 Feb 13;48(3):409–415. doi: 10.1016/0092-8674(87)90192-9. [DOI] [PubMed] [Google Scholar]
  9. Danielpour D., Sporn M. B. Differential inhibition of transforming growth factor beta 1 and beta 2 activity by alpha 2-macroglobulin. J Biol Chem. 1990 Apr 25;265(12):6973–6977. [PubMed] [Google Scholar]
  10. Fontana A., Frei K., Bodmer S., Hofer E., Schreier M. H., Palladino M. A., Jr, Zinkernagel R. M. Transforming growth factor-beta inhibits the generation of cytotoxic T cells in virus-infected mice. J Immunol. 1989 Nov 15;143(10):3230–3234. [PubMed] [Google Scholar]
  11. Fontana A., Hengartner H., de Tribolet N., Weber E. Glioblastoma cells release interleukin 1 and factors inhibiting interleukin 2-mediated effects. J Immunol. 1984 Apr;132(4):1837–1844. [PubMed] [Google Scholar]
  12. Gentry L. E., Lioubin M. N., Purchio A. F., Marquardt H. Molecular events in the processing of recombinant type 1 pre-pro-transforming growth factor beta to the mature polypeptide. Mol Cell Biol. 1988 Oct;8(10):4162–4168. doi: 10.1128/mcb.8.10.4162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gentry L. E., Nash B. W. The pro domain of pre-pro-transforming growth factor beta 1 when independently expressed is a functional binding protein for the mature growth factor. Biochemistry. 1990 Jul 24;29(29):6851–6857. doi: 10.1021/bi00481a014. [DOI] [PubMed] [Google Scholar]
  14. Gentry L. E., Webb N. R., Lim G. J., Brunner A. M., Ranchalis J. E., Twardzik D. R., Lioubin M. N., Marquardt H., Purchio A. F. Type 1 transforming growth factor beta: amplified expression and secretion of mature and precursor polypeptides in Chinese hamster ovary cells. Mol Cell Biol. 1987 Oct;7(10):3418–3427. doi: 10.1128/mcb.7.10.3418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grotendorst G. R., Smale G., Pencev D. Production of transforming growth factor beta by human peripheral blood monocytes and neutrophils. J Cell Physiol. 1989 Aug;140(2):396–402. doi: 10.1002/jcp.1041400226. [DOI] [PubMed] [Google Scholar]
  16. Huang S. S., O'Grady P., Huang J. S. Human transforming growth factor beta.alpha 2-macroglobulin complex is a latent form of transforming growth factor beta. J Biol Chem. 1988 Jan 25;263(3):1535–1541. [PubMed] [Google Scholar]
  17. Jennings J. C., Mohan S. Heterogeneity of latent transforming growth factor-beta isolated from bone matrix proteins. Endocrinology. 1990 Feb;126(2):1014–1021. doi: 10.1210/endo-126-2-1014. [DOI] [PubMed] [Google Scholar]
  18. Jullien P., Berg T. M., Lawrence D. A. Acidic cellular environments: activation of latent TGF-beta and sensitization of cellular responses to TGF-beta and EGF. Int J Cancer. 1989 May 15;43(5):886–891. doi: 10.1002/ijc.2910430525. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Kasafírek E., Fric P., Malis F. The significance of the N-acyl residue of L-alanyl-L-alanyl-L-alanine p-nitroanilide for cleavage by pancreatic elastase. FEBS Lett. 1974 Apr 1;40(2):353–356. doi: 10.1016/0014-5793(74)80262-0. [DOI] [PubMed] [Google Scholar]
  21. Kehrl J. H., Wakefield L. M., Roberts A. B., Jakowlew S., Alvarez-Mon M., Derynck R., Sporn M. B., Fauci A. S. Production of transforming growth factor beta by human T lymphocytes and its potential role in the regulation of T cell growth. J Exp Med. 1986 May 1;163(5):1037–1050. doi: 10.1084/jem.163.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kovacina K. S., Steele-Perkins G., Purchio A. F., Lioubin M., Miyazono K., Heldin C. H., Roth R. A. Interactions of recombinant and platelet transforming growth factor-beta 1 precursor with the insulin-like growth factor II/mannose 6-phosphate receptor. Biochem Biophys Res Commun. 1989 Apr 14;160(1):393–403. doi: 10.1016/0006-291x(89)91669-0. [DOI] [PubMed] [Google Scholar]
  23. LaMarre J., Wollenberg G. K., Gauldie J., Hayes M. A. Alpha 2-macroglobulin and serum preferentially counteract the mitoinhibitory effect of transforming growth factor-beta 2 in rat hepatocytes. Lab Invest. 1990 May;62(5):545–551. [PubMed] [Google Scholar]
  24. Lawrence D. A., Pircher R., Jullien P. Conversion of a high molecular weight latent beta-TGF from chicken embryo fibroblasts into a low molecular weight active beta-TGF under acidic conditions. Biochem Biophys Res Commun. 1985 Dec 31;133(3):1026–1034. doi: 10.1016/0006-291x(85)91239-2. [DOI] [PubMed] [Google Scholar]
  25. Lucas C., Bald L. N., Fendly B. M., Mora-Worms M., Figari I. S., Patzer E. J., Palladino M. A. The autocrine production of transforming growth factor-beta 1 during lymphocyte activation. A study with a monoclonal antibody-based ELISA. J Immunol. 1990 Sep 1;145(5):1415–1422. [PubMed] [Google Scholar]
  26. 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]
  27. Lyons R. M., Keski-Oja J., Moses H. L. Proteolytic activation of latent transforming growth factor-beta from fibroblast-conditioned medium. J Cell Biol. 1988 May;106(5):1659–1665. doi: 10.1083/jcb.106.5.1659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lyons R. M., Moses H. L. Transforming growth factors and the regulation of cell proliferation. Eur J Biochem. 1990 Feb 14;187(3):467–473. doi: 10.1111/j.1432-1033.1990.tb15327.x. [DOI] [PubMed] [Google Scholar]
  29. Madisen L., Farrand A. L., Lioubin M. N., Marzowski J., Knox L. B., Webb N. R., Lim J., Purchio A. F. Expression and characterization of recombinant TGF-beta 2 proteins produced in mammalian cells. DNA. 1989 Apr;8(3):205–212. doi: 10.1089/dna.1.1989.8.205. [DOI] [PubMed] [Google Scholar]
  30. Massagué J., Like B. Cellular receptors for type beta transforming growth factor. Ligand binding and affinity labeling in human and rodent cell lines. J Biol Chem. 1985 Mar 10;260(5):2636–2645. [PubMed] [Google Scholar]
  31. McPherson J. M., Sawamura S. J., Ogawa Y., Dineley K., Carrillo P., Piez K. A. The growth inhibitor of African green monkey (BSC-1) cells is transforming growth factors beta 1 and beta 2. Biochemistry. 1989 Apr 18;28(8):3442–3447. doi: 10.1021/bi00434a044. [DOI] [PubMed] [Google Scholar]
  32. Miyazono K., Heldin C. H. Role for carbohydrate structures in TGF-beta 1 latency. Nature. 1989 Mar 9;338(6211):158–160. doi: 10.1038/338158a0. [DOI] [PubMed] [Google Scholar]
  33. Miyazono K., Hellman U., Wernstedt C., Heldin C. H. Latent high molecular weight complex of transforming growth factor beta 1. Purification from human platelets and structural characterization. J Biol Chem. 1988 May 5;263(13):6407–6415. [PubMed] [Google Scholar]
  34. O'Connor-McCourt M. D., Wakefield L. M. Latent transforming growth factor-beta in serum. A specific complex with alpha 2-macroglobulin. J Biol Chem. 1987 Oct 15;262(29):14090–14099. [PubMed] [Google Scholar]
  35. Okada F., Yamaguchi K., Ichihara A., Nakamura T. Purification and structural analysis of a latent form of transforming growth factor-beta from rat platelets. J Biochem. 1989 Aug;106(2):304–310. doi: 10.1093/oxfordjournals.jbchem.a122849. [DOI] [PubMed] [Google Scholar]
  36. Oreffo R. O., Mundy G. R., Seyedin S. M., Bonewald L. F. Activation of the bone-derived latent TGF beta complex by isolated osteoclasts. Biochem Biophys Res Commun. 1989 Feb 15;158(3):817–823. doi: 10.1016/0006-291x(89)92795-2. [DOI] [PubMed] [Google Scholar]
  37. Piao Y. F., Ichijo H., Miyagawa K., Ohashi H., Takaku F., Miyazono K. Latent form of transforming growth factor-beta 1 acts as a potent growth inhibitor on a human erythroleukemia cell line. Biochem Biophys Res Commun. 1990 Feb 28;167(1):27–32. doi: 10.1016/0006-291x(90)91725-8. [DOI] [PubMed] [Google Scholar]
  38. Pircher R., Jullien P., Lawrence D. A. Beta-transforming growth factor is stored in human blood platelets as a latent high molecular weight complex. Biochem Biophys Res Commun. 1986 Apr 14;136(1):30–37. doi: 10.1016/0006-291x(86)90872-7. [DOI] [PubMed] [Google Scholar]
  39. Purchio A. F., Cooper J. A., Brunner A. M., Lioubin M. N., Gentry L. E., Kovacina K. S., Roth R. A., Marquardt H. Identification of mannose 6-phosphate in two asparagine-linked sugar chains of recombinant transforming growth factor-beta 1 precursor. J Biol Chem. 1988 Oct 5;263(28):14211–14215. [PubMed] [Google Scholar]
  40. Sato Y., Rifkin D. B. Inhibition of endothelial cell movement by pericytes and smooth muscle cells: activation of a latent transforming growth factor-beta 1-like molecule by plasmin during co-culture. J Cell Biol. 1989 Jul;109(1):309–315. doi: 10.1083/jcb.109.1.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sato Y., Tsuboi R., Lyons R., Moses H., Rifkin D. B. Characterization of the activation of latent TGF-beta by co-cultures of endothelial cells and pericytes or smooth muscle cells: a self-regulating system. J Cell Biol. 1990 Aug;111(2):757–763. doi: 10.1083/jcb.111.2.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sha X., Brunner A. M., Purchio A. F., Gentry L. E. Transforming growth factor beta 1: importance of glycosylation and acidic proteases for processing and secretion. Mol Endocrinol. 1989 Jul;3(7):1090–1098. doi: 10.1210/mend-3-7-1090. [DOI] [PubMed] [Google Scholar]
  43. Siepl C., Bodmer S., Frei K., MacDonald H. R., De Martin R., Hofer E., Fontana A. The glioblastoma-derived T cell suppressor factor/transforming growth factor-beta 2 inhibits T cell growth without affecting the interaction of interleukin 2 with its receptor. Eur J Immunol. 1988 Apr;18(4):593–600. doi: 10.1002/eji.1830180416. [DOI] [PubMed] [Google Scholar]
  44. Somorin O., Tokura S., Nishi N., Noguchi J. The action of trypsin on synthetic chromogenic arginine substrates. J Biochem. 1979 Jan;85(1):157–162. doi: 10.1093/oxfordjournals.jbchem.a132305. [DOI] [PubMed] [Google Scholar]
  45. Studer A., de Tribolet N., Diserens A. C., Gaide A. C., Matthieu J. M., Carrel S., Stavrou D. Characterization of four human malignant glioma cell lines. Acta Neuropathol. 1985;66(3):208–217. doi: 10.1007/BF00688585. [DOI] [PubMed] [Google Scholar]
  46. Thotakura N. R., Bahl O. P. Enzymatic deglycosylation of glycoproteins. Methods Enzymol. 1987;138:350–359. doi: 10.1016/0076-6879(87)38030-9. [DOI] [PubMed] [Google Scholar]
  47. Tucker R. F., Branum E. L., Shipley G. D., Ryan R. J., Moses H. L. Specific binding to cultured cells of 125I-labeled type beta transforming growth factor from human platelets. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6757–6761. doi: 10.1073/pnas.81.21.6757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wakefield L. M., Smith D. M., Broz S., Jackson M., Levinson A. D., Sporn M. B. Recombinant TGF-beta 1 is synthesized as a two-component latent complex that shares some structural features with the native platelet latent TGF-beta 1 complex. Growth Factors. 1989;1(3):203–218. doi: 10.3109/08977198908997997. [DOI] [PubMed] [Google Scholar]
  49. Wakefield L. M., Smith D. M., Flanders K. C., Sporn M. B. Latent transforming growth factor-beta from human platelets. A high molecular weight complex containing precursor sequences. J Biol Chem. 1988 Jun 5;263(16):7646–7654. [PubMed] [Google Scholar]
  50. Wrann M., Bodmer S., de Martin R., Siepl C., Hofer-Warbinek R., Frei K., Hofer E., Fontana A. T cell suppressor factor from human glioblastoma cells is a 12.5-kd protein closely related to transforming growth factor-beta. EMBO J. 1987 Jun;6(6):1633–1636. doi: 10.1002/j.1460-2075.1987.tb02411.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. de Martin R., Haendler B., Hofer-Warbinek R., Gaugitsch H., Wrann M., Schlüsener H., Seifert J. M., Bodmer S., Fontana A., Hofer E. Complementary DNA for human glioblastoma-derived T cell suppressor factor, a novel member of the transforming growth factor-beta gene family. EMBO J. 1987 Dec 1;6(12):3673–3677. doi: 10.1002/j.1460-2075.1987.tb02700.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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