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  • Secreted glycoproteins of h...
    DENNIS, M. V; WATSON, C. B; HEIFETZ, A

    Journal of cell science 67
    Journal Article

    Human kidney tumour cells in culture incorporated 3Hglucosamine and 35SO4 into glycoprotein products, which were secreted into the culture medium. The effects of sodium butyrate, a known differentiation-inducing agent, on the production of these sulphated glycoproteins were studied. Cells were cultured in the absence or presence of butyrate (2 mM) in serum-containing medium, for various times, and the labelled glycoproteins were partially purified by DEAE-cellulose chromatography. Treatment of these cells with butyrate resulted in an increase in the synthesis of secreted 3Hglucosamine- and 35SO4-labelled glycoproteins over several days of culture. This same increase in levels of 35SO4 incorporation was not observed with B16 melanoma cells. Sodium dodecyl sulphate (SDS)/polyacrylamide gel electrophoresis revealed that five major glycoproteins labelled with 3Hglucosamine also were labelled with 35SO4. The major secreted glycoproteins from cells cultured in the absence or presence of butyrate over a 3-day period were similar by SDS/polyacrylamide gel electrophoresis. Analyses of Pronase-derived glycopeptides indicated that these secreted 3H/35S-labelled glycoproteins contained sulphated oligosaccharides with terminal sialic acid---Gal---GlcNAc residues similar to glycoproteins secreted by vascular endothelial cells.