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31 results on '"Fukuta, Kazuhiro"'

26. Characterization of Fibroblast Growth Factor-6 Expressed by Chinese Hamster Ovary Cells as a Glycosylated Mitogen for Human Vascular Endothelial Cells

Author

Asada, Masahiro, primary, Yoneda, Atsuko, additional, Oda, Yuko, additional, Ota, Keiko, additional, Ozawa, Kazuo, additional, Fukuta, Kazuhiro, additional, Omae, Fumio, additional, Asanagi, Mineko, additional, Orikasa, Noriko, additional, Suzuki, Masashi, additional, Oka, Syuichi, additional, Makino, Tadashi, additional, and Imamura, Toru, additional

Published
1999
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30. Remodeling of sugar chain structures of human interferon-γ.

Author

Fukuta, Kazuhiro, Abe, Reiko, Yokomatsu, Tomoko, Kono, Naoko, Asanagi, Mineko, Omae, Fumio, Minowa, Mari Toba, Takeuchi, Makoto, and Makino, Tadashi

Abstract

Natural human interferon (IFN)-γ has mainly biantennary complex-type sugar chains and scarcely has multiantennary structures. We attempted to remodel the sugar chain structures using IFN-γ as a model glycoprotein. To obtain the branching glycoforms of IFN-γ, we introduced the genes for GnT-IV (UDP-N-acetylglucosamine:α-1,3-D-mannoside β-1,4-N-acetylglucosaminyltransferase) and/or GnT-V (UDP-N-acetylglucosamine:α-1,6-D-mannoside β-1,6-N-acetylglucosaminyltransferase) into Chinese hamster ovary (CHO) cells producing human IFN-γ. The parental CHO cells produced IFN-γ with biantennary sugar chains mainly. When the GnT-IV activity was increased, triantennary sugar chains with a branch produced by GnT-IV increased up to 66.9% of the total sugar chains. When the GnT-V activity was increased, triantennary sugar chains with a corresponding branch increased up to 55.7% of the total sugar chains. When the GnT-IV and -V activities were increased at a time, tetraantennary sugar chains increased up to 56.2% of the total sugar chains. The proportion of these multiantennary sugar chains corresponded to the intracellular activities of GnT-IV and -V. What is more, lectin blot and flow cytometric analysis indicated that the multi-branch structure of the sugar chains was increased not only on IFN-γ, one of the secretory glycoproteins, but also on almost CHO cellular proteins by introducing either or both of the GnT genes. The results suggest that the branching structure of sugar chains of glycoproteins could be controlled by cellular GnT-IV and GnT-V activities. This technology can produce glycoforms out of natural occurrence, which should enlarge the potency of glycoprotein therapeutics. [ABSTRACT FROM PUBLISHER]

Published
2000
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31. Purification and characterization of glycerate kinase from a serine-producing methylotroph, <em>Hyphomicrobium methlovorum</em> GM2.

Author

Yoshida, Toyokazu, Fukuta, Kazuhiro, Mitsunaga, Toshio, Yamada, Hideaki, and Izumi, Yoshikazu

Subjects
*ENZYMES, *SERINE, *METHYLOTROPHIC microorganisms, *BIOTECHNOLOGY, *BIOCHEMISTRY
Abstract

The glycerate kinase of a serine-producing methylotroph, Hyphomicrobium methylovorum GM2, was purified to complete homogeneity and characterized, the first time for an enzyme from a methylotroph. The enzyme was a monomer with a molecular mass about 41-52 kDa. The enzyme was stable against heating at 35°C for 30 min at pH values over 6-10. Maximum activity was observed at pH 8.0 and around 50°C. The Km values for D-glycerate and ATP were 0.13 mM and 0.13 mM, respectively. The enzyme showed high specificity for D-glycerate, and was activated by potassium and ammonium ions. The reaction product of the enzyme was identified as 2-phosphoglycerate. [ABSTRACT FROM AUTHOR]

Published
1992
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