Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Oct 2;131(2):539–549. doi: 10.1083/jcb.131.2.539

Dual role for the latent transforming growth factor-beta binding protein in storage of latent TGF-beta in the extracellular matrix and as a structural matrix protein

PMCID: PMC2199978  PMID: 7593177

Abstract

The role of the latent TGF-beta binding protein (LTBP) is unclear. In cultures of fetal rat calvarial cells, which form mineralized bonelike nodules, both LTBP and the TGF-beta 1 precursor localized to large fibrillar structures in the extracellular matrix. The appearance of these fibrillar structures preceded the appearance of type I collagen fibers. Plasmin treatment abolished the fibrillar staining pattern for LTBP and released a complex containing both LTBP and TGF-beta. Antibodies and antisense oligonucleotides against LTBP inhibited the formation of mineralized bonelike nodules in long-term fetal rat calvarial cultures. Immunohistochemistry of fetal and adult rat bone confirmed a fibrillar staining pattern for LTBP in vivo. These findings, together with the known homology of LTBP to the fibrillin family of proteins, suggest a novel function for LTBP, in addition to its role in matrix storage of latent TGF-beta, as a structural matrix protein that may play a role in bone formation.

Full Text

The Full Text of this article is available as a PDF (7.3 MB).

Selected References

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

  1. Bascom C. C., Wolfshohl J. R., Coffey R. J., Jr, Madisen L., Webb N. R., Purchio A. R., Derynck R., Moses H. L. Complex regulation of transforming growth factor beta 1, beta 2, and beta 3 mRNA expression in mouse fibroblasts and keratinocytes by transforming growth factors beta 1 and beta 2. Mol Cell Biol. 1989 Dec;9(12):5508–5515. doi: 10.1128/mcb.9.12.5508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bellows C. G., Aubin J. E., Heersche J. N., Antosz M. E. Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int. 1986 Mar;38(3):143–154. doi: 10.1007/BF02556874. [DOI] [PubMed] [Google Scholar]
  3. Bonewald L. F., Dallas S. L. Role of active and latent transforming growth factor beta in bone formation. J Cell Biochem. 1994 Jul;55(3):350–357. doi: 10.1002/jcb.240550312. [DOI] [PubMed] [Google Scholar]
  4. Bonewald L. F., Wakefield L., Oreffo R. O., Escobedo A., Twardzik D. R., Mundy G. R. Latent forms of transforming growth factor-beta (TGF beta) derived from bone cultures: identification of a naturally occurring 100-kDa complex with similarity to recombinant latent TGF beta. Mol Endocrinol. 1991 Jun;5(6):741–751. doi: 10.1210/mend-5-6-741. [DOI] [PubMed] [Google Scholar]
  5. Border W. A., Noble N. A., Yamamoto T., Harper J. R., Yamaguchi Y. u., Pierschbacher M. D., Ruoslahti E. Natural inhibitor of transforming growth factor-beta protects against scarring in experimental kidney disease. Nature. 1992 Nov 26;360(6402):361–364. doi: 10.1038/360361a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Centrella M., Horowitz M. C., Wozney J. M., McCarthy T. L. Transforming growth factor-beta gene family members and bone. Endocr Rev. 1994 Feb;15(1):27–39. doi: 10.1210/edrv-15-1-27. [DOI] [PubMed] [Google Scholar]
  8. Davis E. C. Immunolocalization of microfibril and microfibril-associated proteins in the subendothelial matrix of the developing mouse aorta. J Cell Sci. 1994 Mar;107(Pt 3):727–736. doi: 10.1242/jcs.107.3.727. [DOI] [PubMed] [Google Scholar]
  9. Dietz H. C., Cutting G. R., Pyeritz R. E., Maslen C. L., Sakai L. Y., Corson G. M., Puffenberger E. G., Hamosh A., Nanthakumar E. J., Curristin S. M. Marfan syndrome caused by a recurrent de novo missense mutation in the fibrillin gene. Nature. 1991 Jul 25;352(6333):337–339. doi: 10.1038/352337a0. [DOI] [PubMed] [Google Scholar]
  10. Dworetzky S. I., Fey E. G., Penman S., Lian J. B., Stein J. L., Stein G. S. Progressive changes in the protein composition of the nuclear matrix during rat osteoblast differentiation. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4605–4609. doi: 10.1073/pnas.87.12.4605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Edwards D. R., Murphy G., Reynolds J. J., Whitham S. E., Docherty A. J., Angel P., Heath J. K. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor. EMBO J. 1987 Jul;6(7):1899–1904. doi: 10.1002/j.1460-2075.1987.tb02449.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Flaumenhaft R., Abe M., Sato Y., Miyazono K., Harpel J., Heldin C. H., Rifkin D. B. Role of the latent TGF-beta binding protein in the activation of latent TGF-beta by co-cultures of endothelial and smooth muscle cells. J Cell Biol. 1993 Feb;120(4):995–1002. doi: 10.1083/jcb.120.4.995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Gibson M. A., Kumaratilake J. S., Cleary E. G. The protein components of the 12-nanometer microfibrils of elastic and nonelastic tissues. J Biol Chem. 1989 Mar 15;264(8):4590–4598. [PubMed] [Google Scholar]
  15. Hamilton J. A., Lingelbach S., Partridge N. C., Martin T. J. Regulation of plasminogen activator production by bone-resorbing hormones in normal and malignant osteoblasts. Endocrinology. 1985 Jun;116(6):2186–2191. doi: 10.1210/endo-116-6-2186. [DOI] [PubMed] [Google Scholar]
  16. Harris S. E., Bonewald L. F., Harris M. A., Sabatini M., Dallas S., Feng J. Q., Ghosh-Choudhury N., Wozney J., Mundy G. R. Effects of transforming growth factor beta on bone nodule formation and expression of bone morphogenetic protein 2, osteocalcin, osteopontin, alkaline phosphatase, and type I collagen mRNA in long-term cultures of fetal rat calvarial osteoblasts. J Bone Miner Res. 1994 Jun;9(6):855–863. doi: 10.1002/jbmr.5650090611. [DOI] [PubMed] [Google Scholar]
  17. Ignotz R. A., Massagué J. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem. 1986 Mar 25;261(9):4337–4345. [PubMed] [Google Scholar]
  18. 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]
  19. Kim S. J., Angel P., Lafyatis R., Hattori K., Kim K. Y., Sporn M. B., Karin M., Roberts A. B. Autoinduction of transforming growth factor beta 1 is mediated by the AP-1 complex. Mol Cell Biol. 1990 Apr;10(4):1492–1497. doi: 10.1128/mcb.10.4.1492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kim S. J., Glick A., Sporn M. B., Roberts A. B. Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem. 1989 Jan 5;264(1):402–408. [PubMed] [Google Scholar]
  21. Kubota S., Fridman R., Yamada Y. Transforming growth factor-beta suppresses the invasiveness of human fibrosarcoma cells in vitro by increasing expression of tissue inhibitor of metalloprotease. Biochem Biophys Res Commun. 1991 Apr 15;176(1):129–136. doi: 10.1016/0006-291x(91)90899-i. [DOI] [PubMed] [Google Scholar]
  22. Laiho M., Saksela O., Keski-Oja J. Transforming growth factor-beta induction of type-1 plasminogen activator inhibitor. Pericellular deposition and sensitivity to exogenous urokinase. J Biol Chem. 1987 Dec 25;262(36):17467–17474. [PubMed] [Google Scholar]
  23. Lee B., Godfrey M., Vitale E., Hori H., Mattei M. G., Sarfarazi M., Tsipouras P., Ramirez F., Hollister D. W. Linkage of Marfan syndrome and a phenotypically related disorder to two different fibrillin genes. Nature. 1991 Jul 25;352(6333):330–334. doi: 10.1038/352330a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Lönnqvist L., Child A., Kainulainen K., Davidson R., Puhakka L., Peltonen L. A novel mutation of the fibrillin gene causing ectopia lentis. Genomics. 1994 Feb;19(3):573–576. doi: 10.1006/geno.1994.1110. [DOI] [PubMed] [Google Scholar]
  27. Maddox B. K., Sakai L. Y., Keene D. R., Glanville R. W. Connective tissue microfibrils. Isolation and characterization of three large pepsin-resistant domains of fibrillin. J Biol Chem. 1989 Dec 15;264(35):21381–21385. [PubMed] [Google Scholar]
  28. Marigo V., Volpin D., Vitale G., Bressan G. M. Identification of a TGF-beta responsive element in the human elastin promoter. Biochem Biophys Res Commun. 1994 Mar 15;199(2):1049–1056. doi: 10.1006/bbrc.1994.1335. [DOI] [PubMed] [Google Scholar]
  29. Maslen C. L., Corson G. M., Maddox B. K., Glanville R. W., Sakai L. Y. Partial sequence of a candidate gene for the Marfan syndrome. Nature. 1991 Jul 25;352(6333):334–337. doi: 10.1038/352334a0. [DOI] [PubMed] [Google Scholar]
  30. McKay N. G., Khong T. F., Haites N. E., Power D. A. The effect of transforming growth factor beta 1 on mesangial cell fibronectin synthesis: increased incorporation into the extracellular matrix and reduced pI but no effect on alternative splicing. Exp Mol Pathol. 1993 Dec;59(3):211–224. doi: 10.1006/exmp.1993.1040. [DOI] [PubMed] [Google Scholar]
  31. Milewicz D. M., Pyeritz R. E., Crawford E. S., Byers P. H. Marfan syndrome: defective synthesis, secretion, and extracellular matrix formation of fibrillin by cultured dermal fibroblasts. J Clin Invest. 1992 Jan;89(1):79–86. doi: 10.1172/JCI115589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Miyazono K., Olofsson A., Colosetti P., Heldin C. H. A role of the latent TGF-beta 1-binding protein in the assembly and secretion of TGF-beta 1. EMBO J. 1991 May;10(5):1091–1101. doi: 10.1002/j.1460-2075.1991.tb08049.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mizoi T., Ohtani H., Miyazono K., Miyazawa M., Matsuno S., Nagura H. Immunoelectron microscopic localization of transforming growth factor beta 1 and latent transforming growth factor beta 1 binding protein in human gastrointestinal carcinomas: qualitative difference between cancer cells and stromal cells. Cancer Res. 1993 Jan 1;53(1):183–190. [PubMed] [Google Scholar]
  35. Morén A., Olofsson A., Stenman G., Sahlin P., Kanzaki T., Claesson-Welsh L., ten Dijke P., Miyazono K., Heldin C. H. Identification and characterization of LTBP-2, a novel latent transforming growth factor-beta-binding protein. J Biol Chem. 1994 Dec 23;269(51):32469–32478. [PubMed] [Google Scholar]
  36. Penttinen R. P., Kobayashi S., Bornstein P. Transforming growth factor beta increases mRNA for matrix proteins both in the presence and in the absence of changes in mRNA stability. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1105–1108. doi: 10.1073/pnas.85.4.1105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pfeilschifter J., Erdmann J., Schmidt W., Naumann A., Minne H. W., Ziegler R. Differential regulation of plasminogen activator and plasminogen activator inhibitor by osteotropic factors in primary cultures of mature osteoblasts and osteoblast precursors. Endocrinology. 1990 Feb;126(2):703–711. doi: 10.1210/endo-126-2-703. [DOI] [PubMed] [Google Scholar]
  38. Roberts C. J., Birkenmeier T. M., McQuillan J. J., Akiyama S. K., Yamada S. S., Chen W. T., Yamada K. M., McDonald J. A. Transforming growth factor beta stimulates the expression of fibronectin and of both subunits of the human fibronectin receptor by cultured human lung fibroblasts. J Biol Chem. 1988 Apr 5;263(10):4586–4592. [PubMed] [Google Scholar]
  39. Rosenbloom J., Abrams W. R., Mecham R. Extracellular matrix 4: the elastic fiber. FASEB J. 1993 Oct;7(13):1208–1218. [PubMed] [Google Scholar]
  40. Rossi P., Karsenty G., Roberts A. B., Roche N. S., Sporn M. B., de Crombrugghe B. A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-beta. Cell. 1988 Feb 12;52(3):405–414. doi: 10.1016/s0092-8674(88)80033-3. [DOI] [PubMed] [Google Scholar]
  41. Sakai L. Y., Keene D. R., Engvall E. Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils. J Cell Biol. 1986 Dec;103(6 Pt 1):2499–2509. doi: 10.1083/jcb.103.6.2499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sakai L. Y., Keene D. R., Glanville R. W., Bächinger H. P. Purification and partial characterization of fibrillin, a cysteine-rich structural component of connective tissue microfibrils. J Biol Chem. 1991 Aug 5;266(22):14763–14770. [PubMed] [Google Scholar]
  43. Sato Y., Okada F., Abe M., Seguchi T., Kuwano M., Sato S., Furuya A., Hanai N., Tamaoki T. The mechanism for the activation of latent TGF-beta during co-culture of endothelial cells and smooth muscle cells: cell-type specific targeting of latent TGF-beta to smooth muscle cells. J Cell Biol. 1993 Dec;123(5):1249–1254. doi: 10.1083/jcb.123.5.1249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. Skerry T. M., Bitensky L., Chayen J., Lanyon L. E. Early strain-related changes in enzyme activity in osteocytes following bone loading in vivo. J Bone Miner Res. 1989 Oct;4(5):783–788. doi: 10.1002/jbmr.5650040519. [DOI] [PubMed] [Google Scholar]
  46. Sporn M. B., Roberts A. B. Transforming growth factor-beta: recent progress and new challenges. J Cell Biol. 1992 Dec;119(5):1017–1021. doi: 10.1083/jcb.119.5.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Taipale J., Miyazono K., Heldin C. H., Keski-Oja J. Latent transforming growth factor-beta 1 associates to fibroblast extracellular matrix via latent TGF-beta binding protein. J Cell Biol. 1994 Jan;124(1-2):171–181. doi: 10.1083/jcb.124.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Tomooka S., Border W. A., Marshall B. C., Noble N. A. Glomerular matrix accumulation is linked to inhibition of the plasmin protease system. Kidney Int. 1992 Dec;42(6):1462–1469. doi: 10.1038/ki.1992.442. [DOI] [PubMed] [Google Scholar]
  49. Tsuji T., Okada F., Yamaguchi K., Nakamura T. Molecular cloning of the large subunit of transforming growth factor type beta masking protein and expression of the mRNA in various rat tissues. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8835–8839. doi: 10.1073/pnas.87.22.8835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Van Obberghen-Schilling E., Roche N. S., Flanders K. C., Sporn M. B., Roberts A. B. Transforming growth factor beta 1 positively regulates its own expression in normal and transformed cells. J Biol Chem. 1988 Jun 5;263(16):7741–7746. [PubMed] [Google Scholar]
  51. 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]
  52. Yee J. A., Yan L., Dominguez J. C., Allan E. H., Martin T. J. Plasminogen-dependent activation of latent transforming growth factor beta (TGF beta) by growing cultures of osteoblast-like cells. J Cell Physiol. 1993 Dec;157(3):528–534. doi: 10.1002/jcp.1041570312. [DOI] [PubMed] [Google Scholar]
  53. Zhang H., Apfelroth S. D., Hu W., Davis E. C., Sanguineti C., Bonadio J., Mecham R. P., Ramirez F. Structure and expression of fibrillin-2, a novel microfibrillar component preferentially located in elastic matrices. J Cell Biol. 1994 Mar;124(5):855–863. doi: 10.1083/jcb.124.5.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Zimmermann D. R., Dours-Zimmermann M. T., Schubert M., Bruckner-Tuderman L. Versican is expressed in the proliferating zone in the epidermis and in association with the elastic network of the dermis. J Cell Biol. 1994 Mar;124(5):817–825. doi: 10.1083/jcb.124.5.817. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES