Abstract
Human skin fibroblasts were incubated with a fluorogenic xyloside, 4-methylumbelliferyl-β-D-xyloside (Xyl-MU), in the presence or absence of tunicamycin. The xyloside-initiated glycosaminoglycans (GAG-MUs) were isolated from the culture medium, and their structures characterized. When the cells were incubated with Xyl-MU in the presence of 0.2 μg ml−1 tunicamycin, the synthesis of GAG-MU was increased about three fold, compared with the control value in the absence of tunicamycin (cells exposed to Xyl-MU alone). The structures of GAG-MUs synthesized in the presence or absence of tunicamycin were compared by HPLC analysis using gel-filtration and ion-exchange columns, enzymatic digestion, and unsaturated disaccharide composition analysis. The data indicated that cells incubated with tunicamycin produced more undersulfated and shorter GAG-MUs than cells without tynicamycin. These results suggest that tunicamycin inhibits the elongation and sulfation of glycosaminoglycan (GAG) chains and that, as a result, GAG-MUs with shorter chains and undersulfated residues, but possessing a large number of GAG chains, are synthesized in the presence of tunicamycin.
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References
Okayama M, Kimata K, Suzuki S (1973) J Biochem 74: 1069–73.
Schwartz NB, Galligani L, Ho P-L, Dorfman A (1974) Proc Natl Acad Sci USA 71: 4047–51.
Fukunaga Y, Sobue M, Suzuki N, Kushida H, Suzuki S, Suzuki S (1975) Biochim Biophys Acta 381: 443–7.
Robinson HC, Brett MJ, Tralaggan PJ, Lowther DA, Okayama M (1975) Biochem J 148: 25–34.
Kato Y, Kimata K, Ito K, Karasawa K, Suzuki S (1978) J Biol Chem 253: 2784–9.
Kolset SO, Ehlorsson J, Kjellén L, Lindahl U (1986) Biochem J 238: 209–16.
Sobue M, Habuchi H, Ito K, Yonekura H, Oguri K, Sakurai K, Kamohara S, Ueno Y, Noyori R, Suzuki S (1987) Biochem J 241: 591–601.
Lugemwa FN, Esko JD (1991) J Biol Chem 266: 6674–7.
Fransson L-Å, Havsmark B, Sakurai K, Suzuki S (1992) Glycoconjugate J 9: 45–55.
Takagaki K, Nakamura T, Kon A, Tamura S, Endo M (1991) J Biochem 109: 514–19.
Freeze HH, Sampath D, Varki A (1993) J Biol Chem 268: 1618–27.
Takagaki K, Nakamura T, Shibata S, Higuchi T, Endo M (1996) J Biochem 119: 697–702.
Nakamura T, Izumi J, Takagaki K, Shibata S, Kojima K, Kato I, Endo M (1994) Biochem J 304: 731–6.
Shibata S, Takagaki K, Nakamura T, Izumi J, Kojima K, Kato I, Endo M (1995) J Biol Chem 270: 13794–8.
Izumi J, Takagaki K, Nakamura T, Shibata S, Kojima K, Kato I, Endo M (1994) Biochem 116: 524–9.
Takatsuki A, Kohno K, Tamura G (1975) Agric Biol Chem 39: 2089–91.
Tkacz JS, Lampen JO (1975) Biochem Biophys Res Commun 65: 248–57.
Struck DK, Lennarz WJ (1997) J Biol Chem 252: 1007–13.
Hart GW, Lennarz WJ (1978) J Biol Chem 253: 5795–801.
Takagaki K, Kon A, Kawasaki H, Nakamura T, Tamura S, Endo M (1990) J Biol Chem 265: 854–60.
Nakamura T, Takagaki K, Shibata S, Tanaka K, Higuchi H, Endo M (1995) Biochem Biophys Res Commun 208: 470–5.
Saito H, Yamagata T, Suzuki S (1968) J Biol Chem 243: 1536–42.
Linker A, Hovingh P (1975) Methods Enzymol 28: 902–11.
Ohya T, Kaneko Y (1970) Biochim Biophys Acta 198: 607–9.
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Takagaki, K., Tazawa, T., Munakata, H. et al. Characterization of β-D-xyloside-initiated glycosaminoglycan synthesized by human skin fibroblasts in the presence of tunicamycin. Glycoconj J 15, 483–489 (1998). https://doi.org/10.1023/A:1006935003534
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DOI: https://doi.org/10.1023/A:1006935003534