Animal cells and systems

  • 제4권3호
  • /
  • Pages.273-278
  • /
  • 2000
  • /
  • 1976-8354(pISSN)
  • /
  • 2151-2485(eISSN)
한국통합생물학회 (The Korean Society for Integrative Biology)
권호 표지

Expression and Activation of Transforming Growth Factor-Beta 2 in Cultured Bone Cells

  • Lee, Chang-Ho (Department of Biology, College of Natural Sciences, Kangnung National University)
  • 발행 : 2000.09.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Transforming growth factor-$\beta$ (IGF-$\beta$)s are multifunctional small polypeptides synthesized in most cell types. TGF-$\beta$ exerts pivotal effects on both bone formation and resorption. In addition, increasing lines of evidence implicate TGF-$\beta$ as a potential coupling factor between these two processes during bone remodeling. In the present study, the expression form and the activation mechanism of latent-TGF-$\beta$ were investigated using specific antibodies for each isoform. TGF-$\beta$s were observed to be synthesized and accumulated in a large amount in cultured osteoblastic cells. The estimated molecular weights of intracellular TGF-$\beta$2 and -$\beta$3 were 49 and 55 kDa, respectively. Based on proteolytic digestion study and immunofluorescence observation, these precursor forms seemed to be accumulated in distinct intracellular compartments. To examine whether the internal pool of TGF-$\beta$ was possiblely regulated by external signals, their biological activites were examined in a conditioned media of this cell. Although the intact conditioned media did not contain detectable TGF-$\beta$ activity, heat-treatment or acid-activation of the conditioned media revealed significant TGF-$\beta$ activity. Furthermore, in the presence of estrogen, this activity was dramatically diminished. It is known that activation of latent TGF-$\beta$ can be achieved by different chemical and enzymatic treatments, or by incubation with certain cell types. This extracellular activation was suggested as a key step in the regulation of TGF-$\beta$ activity. In addition to these extracellular activation, this study suggests that the synthesis and intracellular processing are important regulation steps for TGF-$\beta$ action. In addition, this regulation Is specific for TGF-$\beta$ type 2, because the change was not observed in TGF-$\beta$3 in osteoblastic cell line.

키워드

참고문헌

  1. Barr PJ (1991) Mammalian subtilisins: the long-sought dibasic processing endoproteases. Cell 66: 1-3 https://doi.org/10.1016/0092-8674(91)90129-M
  2. Bonewald LF, Wakefield L, Oreffo ROC, Escobedo A, Twardzik DR, and Mundy GR (1991) Latent forms of transforming growth $factor-{\beta}(TGF-{\beta})$ derived from bone cultures: identification of naturally occurring 100-kDa complex with similarity to recombinant latent $TGF-{\beta}$. Mol Endocrinol 5: 741-751
  3. Bonewald LF (1999) Regulation and regulatory activities of transforming growth factor-beta. Crit Rev Eukaryot Gene Expr 9: 33-44
  4. Chong H, Vodovotz Y, Cox GW, and Barcellos-Hoff MH (1999) Immunocytochemical localization of latent transforming growth factor-beta1 activation by stimulated macro phages. J Cell Physiol 178: 275-83 https://doi.org/10.1002/(SICI)1097-4652(199903)178:3<275::AID-JCP1>3.0.CO;2-Q
  5. Chu TM and Kawinski E (1998) Plasmin, substilisin-Iike endoproteases, tissue plasminogen activator, and urokinase plasminogen activator are involved in activation of latent TGFbeta 1 in human seminal plasma. Biochem Biophys Res Commun 253: 128-134 https://doi.org/10.1006/bbrc.1998.9760
  6. Crawford SE, Stellmach V, Murphy-Ullrich JE, Ribeiro SM, Lawler J, Hynes RO, Boivin GP, and Bouck N (1998) Thrombospondin-1 is a major activator of TGF-beta1 in vivo. Cell 93: 1159-70 https://doi.org/10.1016/S0092-8674(00)81460-9
  7. Dallas SL, Miyazono K, Skerry TM, Mundy GR, and Bonewald LF (1995) Dual role for the latent transforming growth factor beta binding protein (L TBP) in storage of latent TGF-${\beta}$ in the extracellular matrix and as a structural matrix protein. J Cell Biol 131: 539-549 https://doi.org/10.1083/jcb.131.2.539
  8. Erlebacher A and Derynck R (1996) Increase expression of TGF-beta 2 in ostoblasts results in osteoporosis-like phenotype. J Cell Biol 132: 195-210 https://doi.org/10.1083/jcb.132.1.195
  9. Grainger DJ, Kemp PR, Liu AC, Lawn RM, and Wetcalfe JC (1994) Activation of transforming growth factor-beta is inhibited in transgenic apolipoprotein(a) mice. Nature 370: 460-462 https://doi.org/10.1038/370460a0
  10. Huang SS, Zhou M, Johonson FE, Shiel HS, and Huang JS (1999) An active site of transforming growth factor-beta(1) for growth inhibition and stimulation. J Biol Chem 274: 27754-27758 https://doi.org/10.1074/jbc.274.39.27754
  11. Lioubin M, Madisen L, Roth RA, and Purchio AF (1991) Characterization of latent transforming growth factor-beta2 from monkey kidney cells. Endocrinology 128: 2291-2296
  12. Miyazono K, Thyberg J, and Heldin CH (1992) Retention of the transforming growth factor-beta1 precursor in the Golgi complex in a latent endoglycosidase H-sensitive form. J Biol Chem 267: 5668-5675
  13. Nakayama K (1997) Furin: a mammalian subtilisin/Kex2p-like endoprotease involved in processing of a wide variety of precursor proteins. Biochem J 327: 625-35
  14. Oreffo ROC, Mundy GR, Seyedin S, and Bonewald LF (1989) Activation of the bone derived latent recombinant transforming growth factor-beta complex by isolated osteoclasts. Biochem Biophy Res Commun 158: 817-823 https://doi.org/10.1016/0006-291X(89)92795-2
  15. Roth S, Michel K, and Gressner AM (1998) (Latent) transforming growth factor beta in liver parenchymal cells, its injurydependent release, and paracrine effects on rat hepatic stellate cells. Hepatology 27: 1003-1012 https://doi.org/10.1002/hep.510270416
  16. Roth-Eichhorn S, Khl K, and Gressner AM (1998) Subcellular localization of (latent) transforming growth factor beta and the latent TGF-beta binding protein in rat hepatocytes and hepatic stellate cells. Hepatology 28: 1588-96 https://doi.org/10.1002/hep.510280619
  17. Rowley DR (1992) Glucocorticoid regulation of transforming growth factor-beta activation in urogenital sinus mesenchymal cells. Endocrinology 131: 471-478 https://doi.org/10.1210/en.131.1.471
  18. Sato Y and Rifkin DB (1989) Inhibition of endothelial cell movement by pericytes and smooth muscle cells: activation of a latent transforming growth factor-beta1-like molecular by plasmin during coculture. J Cell Biol 109: 309-315 https://doi.org/10.1083/jcb.109.1.309
  19. Schalch L, Rordorf-Adam C, Dasch JR, and Jungi TW (1991) IgG-stimulated and LPS-stimulated monocytes elaborate transforming growth factor type b (TGF-Il) in active form. Biochem Biophy Res Commun 174: 885-891 https://doi.org/10.1016/0006-291X(91)91500-C
  20. Twardzik DR, Mikovits JA, Ranchalis JE, Purchio AF, Elingsworth L, and Ruscetti FW (1990) Gamma-interferon-induced activation of latent transforming growth factor-beta by human monocytes. Ann N Y Acad Sci 593: 276-284 https://doi.org/10.1111/j.1749-6632.1990.tb16119.x
  21. Wakefield LM, Smith OM, Fladers KC, and Sporn MB (1988) Latent transforming growth factor-beta from human platelet. A high molecular weight complex containing precursor sequences. J Biol Chem 263: 7646-7654
  22. Zhu Y, Oganesian A, Keene DR, and Sandell LJ (1999) Type IIA procollagen containing the cysteine-rich amino pro peptide is deposited in the extracellular matrix of prechondrogenic tissue and binds to TGF-beta1 and BMP-2. J Cell Biol 144: 1069-80 https://doi.org/10.1083/jcb.144.5.1069