caa a22 4500 39755754X CHVBK 20180308164807.0 cr unu---uuuuu 161202e199604 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021298 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021298 Characterization and Biological Significance of Sialylα2-3Galactosylβ1-4Xylosylβl-(4-Methylumbelliferone) Synthesized in Cultured Human Skin Fibroblasts [Elektronische Daten] [Keiichi Takagaki, Toshiya Nakamura, Shigeru Shibata, Tsuyoshi Higuchi, Masahiko Endo] Human skin fibroblasts were incubated in the presence of a fluorogenic xyloside, 4-methyl-umbelliferyl-β-D-xyloside (Xyl-MU), then the cultured medium was recovered, concentrated with a lyophilizer, and dialyzed against distilled water. The structures of the Xyl-MU derivatives purified from the dialyzable fraction were investigated. In addition to established glycosaminoglycans-MU (GAGs-MU), Gal-Gal-Xyl-MU, Gal-Xyl-MU, sul-phate-GlcA-Xyl-MU, GlcA-Xyl-MU, and Xyl-Xyl-MU, which were induced by Xyl-MU, an oligosaccharide having fluorescence was purified using a combination of gel filtration, ion-exchange chromatography and high-performance liquid chromatography, then subjected to carbohydrate composition analysis, enzyme digestion, Smith degradation, 1H-NMR, and ion-spray mass spectrometric analysis. From the data obtained, the oligosaccharide was considered to have the structure SAα2-3Galβ1-4Xylβl-MU. The amount of MU-oligosaccharide in the cell culture increased with time and was dependent on the amount of Xyl-MU added. Its production was also different from that of Gal-Gal-Xyl-MU and Gal-Xyl-MU, which are biosynthetic intermediates of GAG-MU. Addition of CDP, an inhibitor of sialyltransferase, to the cell culture medium increased the secretion of GAG-MU. These results suggest that SA-Gal-Xyl-MU production may be related to the regulation of GAG-MU biosynthesis. © by the Journal of Biochemistry Regular Papers nationallicence cultured human skin fibroblast nationallicence glycosaminoglycan nationallicence 4-methylumbelliferyl-β-D-xyloslde nationallicence SAα2-3Galβl-4Xylβl-(4-methylumbelliferone) nationallicence Takagaki Keiichi Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut Nakamura Toshiya Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut Shibata Shigeru Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut Higuchi Tsuyoshi Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut Endo Masahiko Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut The Journal of Biochemistry Oxford University Press 119/4(1996-04), 697-702 0021-924X 119:4<697 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021298 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021298 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Takagaki Keiichi Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut NATIONALLICENCE 700 1- Nakamura Toshiya Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut NATIONALLICENCE 700 1- Shibata Shigeru Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut NATIONALLICENCE 700 1- Higuchi Tsuyoshi Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut NATIONALLICENCE 700 1- Endo Masahiko Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/4(1996-04), 697-702 0021-924X 119:4<697 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford