Abstract
The human monocytic cell line U-937 has been widely used as a model system for human monocytes. The subclone U-937-B has been adapted to serum-free conditions. This particular U-937 clone and its parent clone U-937-1 were used to investigate the role of the proteoglycan serglycin in human monocytes. For this purpose cells were treated with hexyl-β-D-thioxyloside to abrogate proteoglycan expression. U-937-B cells expressed and secreted exclusively chondroitin sulphate proteoglycans, and after treatment with this xyloside they only expressed and released free chondroitin sulphate chains. Western blotting showed that serglycin core protein was present in conditioned medium of control cells, but absent in medium from xyloside-treated cells. Also, serglycin core protein could be detected in the cell fractions of control cells, but not in the cell fractions from xyloside-treated cells. Furthermore, less proteoglycan-associated proteins could be detected in medium from cells incubated with xyloside, suggesting that the absence of secreted sergycin affects the secretion of such proteins. Cells incubated in the presence of xyloside were analyzed by transmission electron microscopy and shown to contain numerous large empty vesicles. The lack of serglycin, the dominant proteoglycan in U-937 monocyte-like cells, consequently, leads to effects on vesicle formation and secretion of some low molecular weight proteins, suggesting that this particular proteoglycan is of importance for secretory processes in human monocytes.
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References
Abrink, M., Grujic, M., Pejler, G.: Serglycin is essential for maturation of mast cell secretory granule. J. Biol. Chem. 279 J, 40897–40905 (2004)
Belting, M., Borsig, L., Fuster, M.M., Brown, J.R., Persson, L., Fransson, L.A., Esko, J.D.: Tumor attenuation by combined heparan sulfate and polyamine depletion. Proc. Natl. Acad. Sci. USA. J 99, 371–376 (2002)
Bitter, T., Muir, H.M.: A modified uronic acid carbazole reaction. Anal. Biochem. 4, 330–334 (1962)
Esko, J.D., Selleck, S.B.: Order out of chaos: assembly of ligand binding sites in heparan sulfate. Annu. Rev. Biochem. 71, 435–471 (2002)
Grujic, M., Braga, T., Lukinius, A., Eloranta, M.L., Knight, S.D., Pejler, G., Abrink, M.: Serglycin-deficient cytotoxic T lymphocytes display defective secretory granule maturation and granzyme B storage. J. Biol. Chem. 280, 33411–33418 (2005)
Handel, T.M., Johnson, Z., Crown, S.E., Lau, E.K., Proudfoot, A.E.: Regulation of protein function by glycosaminoglycans—as exemplified by chemokines. Annu. Rev. Biochem. 74, 385–410 (2005)
Kjellen, L., Lindahl, U.: Proteoglycans: structures and interactions. Annu. Rev. Biochem. 60, 443–475 (1991)
Kolset, S.O.: Proteoglycans in normal and neoplastic monocytes. Exp. Cell. Res. 168, 318–324 (1987)
Kolset, S.O.: Larsen T, Sulfur-containing macromolecules in cultured monocyte-like cells. Acta Histochem. 84, 67–75 (1988)
Kolset, S.O., Mann, D.M., Uhlin-Hansen, L., Winberg, J.O., Ruoslahti, E.: Serglycin-binding proteins in activated macrophages and platelets. J. Leukoc. Biol. 59, 545–554 (1996)
Kolset, S.O., Prydz, K., Pejler, G.: Intracellular proteoglycans. Biochem. J. 379, 217–227 (2004)
Kolset, S.O., Sakurai, K., Ivhed, I., Overvatn, A., Suzuki, S.: The effect of beta-D-xylosides on the proliferation and proteoglycan biosynthesis of monoblastic U-937 cells. Biochem. J. 265, 637–645 (1990)
Lander, A.D., Selleck, S.B.: The elusive functions of proteoglycans: in vivo veritas. J. Cell. Biol. 148, 227–232 (2000)
Lugemwa, F.N., Esko, J.D.: Estradiol beta-D-xyloside, an efficient primer for heparan sulfate biosynthesis. J. Biol. Chem. 266, 6674–6677 (1991)
Nathan, C.F.: Secretory products of macrophages. J. Clin. Invest. 79, 319–326 (1987)
Niemann, C.U., Cowland, J.B., Klausen, P., Askaa, J., Calafat, J., Borregaard, N.: Localization of serglycin in human neutrophil granulocytes and their precursors. J. Leukoc. Biol. 76, 406–415 (2004)
Opdenakker, G., Van den Steen, P.E., Van Damme, J.: Gelatinase B: a tuner and amplifier of immune functions. Trends Immunol. 22, 571–579 (2001)
Pejler, G., Winberg, J.O., Vuong, T.T., Henningsson, F., Uhlin-Hansen, L., Kimata, K., Kolset, S.O.: Secretion of macrophage urokinase plasminogen activator is dependent on proteoglycans. Eur. J. Biochem. 270, 3971–3980 (2003)
Skutelsky, E., Shoichetman, T., Hammel, I.: An histochemical approach to characterization of anionic constituents in mast cell secretory granules. Histochem. Cell. Biol. 104, 453–458 (1995)
Uhlin-Hansen, L., Eskeland, T., Kolset, S.O.: Modulation of the expression of chondroitin sulfate proteoglycan in stimulated human monocytes. J. Biol. Chem. 264, 14916–14922 (1989)
Uhlin-Hansen, L., Langvoll, D., Wik, T., Kolset, S.O.: Blood platelets stimulate the expression of chondroitin sulfate proteoglycan in human monocytes. Blood. 80, 1058–1065 (1992)
Uhlin-Hansen, L., Wik, T., Kjellen, L., Berg, E., Forsdahl, F., Kolset, S.O.: Proteoglycan metabolism in normal and inflammatory human macrophages. Blood. 82, 2880–2889 (1993)
Vassalli, J.D., Pepper, M.S.: Tumour biology. Membrane proteases in focus. Nature. 370, 14–15 (1994)
Zernichow, L., Abrink, M., Hallgren, J., Grujic, M., Pejler, G., Kolset, S.O.: Serglycin is the major secreted proteoglycan in macrophages and has a role in the regulation of macrophage tumor necrosis factor-alpha secretion in response to lipopolysaccharide. J. Biol. Chem. 281, 26792–26801 (2006)
Zernichow, L., Dalen, K.T., Prydz, K., Winberg, J.O., Kolset, S.O.: Secretion of proteases in serglycin transfected Madin-Darby canine kidney cells. Febs. J. 273, 536–547 (2006)
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This work was supported by grants from the Norwegian Cancer Society. The technical assistance of Aud Øvervatn and Parvin Mahzonni is appreciated.
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Kolset, S.O., Zernichow, L. Serglycin and secretion in human monocytes. Glycoconj J 25, 305–311 (2008). https://doi.org/10.1007/s10719-007-9073-9
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DOI: https://doi.org/10.1007/s10719-007-9073-9