caa a22 4500 397558538 CHVBK 20180308164810.0 cr unu---uuuuu 161202e199611 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021492 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021492 Detection of UDP-D-Xylose:α-D-Xyloside α→3Xylosyltransferase Activity in Human Hepatoma Cell Line HepG2 [Elektronische Daten] [Satsuki Minamida, Kyoko Aoki, Shunji Natsuka, Kaoru Omichi, Koichi Fukase, Shoichi Kusumoto, Sumihiro Hase] We previously reported the detection of novel O-linked sugar chains classified as being of the glucosyl-O-serine type [Hase et al. (1988) J. Biochem. 104, 867-868]. The sugar chains are a disaccharide (Xylα1-3Glc) and a trisaccharide (Xylα1-3Xylα1-3Glc) linked to serine residues in epidermal growth factor-like domains of human and bovine blood coagulation factors. The structures of these sugar chains suggested the presence of an a1→xylosyltransferase for their biosynthesis. We report here on the detection of a1→3xylosyltransferase activity which catalyzes the transfer of xylose to Xylα1-3Glc in the human hepatoma cell line HepG2. We employed pyridylaminated Xylα1-3Glc as a fluorescent acceptor and UDP-D-Xyl as a donor. The reaction product was purified by reversed-phase HPLC, and the structure of the transfer product isolated was confirmed to be pyridylaminated Xylαl-3Xylα1-3Glc by Smith degradation, mass spectrometry, and α- and β-xylosidase digestions. The apparent Km value for pyridylaminated Xylα1-3Glc was 52 mM and for UDP-D-Xyl 0.28 mM. Optimum pH was 7.2. The enzyme was inactivated by addition of EDTA, and its activity was restored by addition of Mn2+ and Mg2+. These results indicate the presence of a novel enzyme which is able to transfer xylose to Xylα1-3Glc, forming Xylα1-3Xylα1-3Glc in human cells. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence α-l nationallicence 3-xylosyltransferase nationallicence fluorogenic acceptor nationallicence human hepatoma HepG2 cell line nationallicence Xylαl-3Xylαl-3Glc nationallicence Minamida Satsuki Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut Aoki Kyoko Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut Natsuka Shunji Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut Omichi Kaoru Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut Fukase Koichi Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut Kusumoto Shoichi Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut Hase Sumihiro Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut The Journal of Biochemistry Oxford University Press 120/5(1996-11), 1002-1006 0021-924X 120:5<1002 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021492 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021492 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Minamida Satsuki Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 700 1- Aoki Kyoko Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 700 1- Natsuka Shunji Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 700 1- Omichi Kaoru Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 700 1- Fukase Koichi Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 700 1- Kusumoto Shoichi Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 700 1- Hase Sumihiro Department of Chemistry, Osaka University College of Science, Toyonaka, Osaka 560 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/5(1996-11), 1002-1006 0021-924X 120:5<1002 1996 120 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