Summary
Collagen-binding heat-shock proteins ofM r 46–47 kDa have been postulated to function as putative molecular chaperones in the biosynthesis of collagen in several species. The rat homologue of this family of heat-shock proteins is called gp46. In the present study, we employed Western blotting and immunohistochemical methods to determine the tissue distribution and cellular localization of gp46 in the thoracic aorta, heart, kidney, liver and lung of eight-day-old Wistar rats. Highest levels of gp46 were detected in the thoracic aorta and lung, followed by the kidney and heart. Gp46 levels were low to undetectable by Western blot analysis in the liver. Immunohistochemistry revealed that gp46 labelling was observed almost exclusively in three distinct cell types: fibroblasts, muscle cells, and some epithelial cells. Gp46 was detected in the fibroblasts of the hepatic triad, in the interstitium of the alveolar wall and in the tunica adventitia of blood vessels in the majority of tissues examined, in atrial and ventricular cardiomyocytes, in vascular smooth muscle cells of the abluminal portion of the tunica media, in parietal epithelial cells and mesangial cells of the glomerulus, in epithelial cells of the distal tubules and collecting ducts in the kidney and clusters of immature renal tubules, in epithelial cells of the bile duct, and in mesodermal cells surrounding the liver. These results demonstrate that gp46 is present in collagen producing cells and cells undergoing rapid growth and development, suggesting that gp46 may play a significant role in these processes.
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References
Andres, D.A., Dickerson, I.M. &Dixon, J.E. (1990) Variants of the carboxyl-terminal KDEL sequence direct intracellular retention.J. Biol. Chem. 265, 5952–5.
Beckmann, R.P., Mizzen, L.A. &Welch, W.J. (1990) Interaction of HSP70 with newly synthesized proteins: implications for HSP protein folding and assembly.Science 248, 850–4.
Cates, G.A. (1987) Differentiation defective mutants of skeletal myoblasts altered in a gelatin-binding glycoprotein.Biochem. Cell Biol. 65, 767–75.
Cates, C.A., Brickenden, A.M. &Sanwal, B.D. (1984) Possible involvement of a cell surface glycoprotein in the differentiation of skeletal myoblasts.J. Biol. Chem. 259, 2646–50.
Clarke, E.P. &Sanwal, B.D. (1992) Cloning of a human collagen-binding protein, and its homology with rat gp46, chick hsp47 and mouse J6 proteins.Biochim. Biophys. Acta 1129, 246–8.
Clarke, E.P., Cates, G.A., Ball, E.H. &Sanwal, B.D. (1991) A collagen-binding protein in the endoplasmic reticulum of myoblasts exhibits relationship with serine protease inhibitors.J. Biol. Chem. 266, 17230–5.
Clarke, E.P., Jain, N., Brickenden, A., Lorimer, I.A. &Sanwal, B.D. (1993) Parallel regulation of procollagen I and colligin, a collagen-binding protein and a member of the serine protease inhibitor family.J. Cell Biol. 121, 193–9.
Craig, E., Kang, P.J. &Boorstein, W. (1990) A review of the role of 70 kDa heat shock proteins in protein translocation across membranes.Antonie van Leeuwenhoek 58, 137–46.
Eghbali, M., Blumenfeld, O.O., Seifter, S., Buttrick, R.M., Leinwand, L.A., Robinson, T.F., Zern, M.A. &Giambrone, M.A. (1989) Localization of types I, III and IV collagen mRNAs in rat heart cells by in situ hybridization.J. Mol. Cell. Cardiol. 21, 103–13.
Graham, R.C. &Karnovsky, M.J. (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique.J. Histochem. Cytochem. 14, 291–302.
Hospelhorn, A.C., Martin, B.M. &Franzblau, C. (1992) Type IV collagen synthesis and accumulation in neonatal rat aortic smooth muscle cell cultures.Matrix 12, 352–61.
Johnson, D., Oppermann, H., Jackson, J. &Levinson, W. (1980) Induction of four proteins in chick embryo cells by sodium arsenite.J. Biol. Chem. 255, 6975–80.
Kurkinen, M., Taylor, A., Garrels, J.I. &Hogan, B.L.M. (1984) Cell surface-associated proteins which bind native type IV collagen or gelatin.J. Biol. Chem. 259, 5915–22.
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–5.
Mays, P.K., Mcanulty, R.J., Campa, J.S. &Laurent, G.J. (1991) Age-related changes in collagen synthesis and degradation in rat tissues.Biochem. J. 276, 307–13.
Miyaishi, O., Sakata, K., Matsuyama, M. &Saga, S. (1992) Distribution of the collagen binding heat-shock protein in chicken tissues.J. Histochem. Cytochem. 40, 1021–9.
Munro, S. &Pelham, H.R.B. (1987) A C-terminal signal prevents secretion of lumenal ER proteins.Cell 48, 899–907.
Nagata, K. &Yamada, K.M. (1986) Phosphorylation and transformation sensitivity of a major collagen-binding protein of fibroblasts.J. Biol. Chem. 261, 7531–6.
Nagata, K., Saga, S. &Yamada, K.M. (1986) A major collagen-binding protein of chick embryo fibroblasts is a novel heat shock protein.J. Cell Biol. 103, 223–9.
Nakai, A., Satoh, M., Hirayoshi, K. &Nagata, K. (1992) Involvement of the stress protein hsp47 in procollagen processing in the endoplasmic reticulum.J. Cell Biol. 117, 903–14.
Nandan, D., Cates, G.A., Ball, E.H. &Sanwal, B.D. (1988) A collagen-binding protein involved in the differentiation of myoblasts recognizes the Arg-Gly-Aso sequence.Exp. Cell Res. 179, 289–97.
Nandan, D., Ball, E.H. &Sanwal, B.D. (1990) Two stress proteins of the endoplasmic reticulum bind denatured collagen.Biochem. Cell Biol. 68, 1057–61.
Pak, B.J., Wigle, D.A., Watson, J.D., Cates, G.A., Brickenden, A.M., Ball, E.H. &Pang, S.C. (1996) Developmental expression of the collagen-binding heat-shock protein gp46 and collagen types I and IV in rat tissues.Biochem. Cell Biol. 74, 179–85.
Pelham, H.R.B. (1986) Speculations on the functions of the major heat shock and glucose-regulated proteins.Cell 46, 959–61.
Peterson, G.L. (1977) A simplification of the protein assay method of Lowryet al. which is more generally applicable.Anal. Biochem. 83, 346–56.
Saga, S., Nagata, K., Chen, W.-T. &Yamada, K.M. (1987) pH-dependent function, purification, and intracellular location of a major collagen-binding glycoprotein.J. Cell Biol. 105, 517–27.
Sauk, J.J., Smith, T., Norris, K. &Ferreira, L. (1994) Hsp47 and the translocation-translocation machinery cooperate in the production of α1(I) chains of type I procollagen.J. Biol. Chem. 269, 3941–6.
Taylor, A., Hogan, B.L.M. &Watt, F.M. (1985) Biosynthesis of EGF receptor, transferrin receptor and colligin by cultured human keratinocytes and the effect of retinoic acid.Exp. Cell Res. 159, 47–54.
Vaillancourt, J.P. &Cates, G.A. (1991) Purification and reconstitution of a collagen-binding heat-shock glycoprotein from L6 myoblasts.Biochem. J. 274, 793–8.
Wang, S.-Y., Larosa, G.J. &Gudas, L.J. (1985) Molecular cloning of gene sequences transcriptionally regulated by retinoic acid and dibutyryl cyclic AMP in cultured mouse teratocarcinoma cells.Dev. Biol. 107, 75–86.
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Pak, B.J., Vanhorne, J.B. & Pang, S.C. Tissue distribution and immunohistochemical localization of the collagen-binding heat-shock protein gp46 in neonatal rats. Histochem J 28, 895–902 (1996). https://doi.org/10.1007/BF02331393
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DOI: https://doi.org/10.1007/BF02331393