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Structural analysis of N-acetylglucosaminyltransferase II, an enzyme essential for complex-type N-glycan biosynthesis

Research Project

Project/Area Number 16H06847
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Applied biochemistry
Research InstitutionShizuoka University

Principal Investigator

Miyazaki Takatsugu  静岡大学, グリーン科学技術研究所, 助教 (30775721)

Research Collaborator PARK Enoch Y.  静岡大学, グリーン科学技術研究所, 教授
KATO Tatsuya  静岡大学, 農学部, 准教授
Project Period (FY) 2016-08-26 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords糖鎖 / 糖転移酵素 / 糖タンパク質 / N結合型糖鎖 / 複合型糖鎖 / カイコ / N結合型糖鎖 / X線結晶構造解析 / N-アセチルグルコサミン
Outline of Final Research Achievements

This study focuses on the structure-function relationship of N-acetylglucosaminyltransferase (GnTII) which is essential for complex-type N-glycan biosynthesis. We obtained recombinant GnTII enzymes from human and Bombyx mori using Escherichia coli and silkworm as hosts, and the enzymes expressed in silkworm showed higher activity than those in E. coli. The substrate specificity and enzymatic activity of B. mori GnTII were almost identical to those of human GnTII. Although B. mori GnTII was more highly N-glycosylated than human GnTII, enzymatic cleavage of N-glycans did not affect their activity and thermostability, indicating that N-glycans which were attached to the enzymes are not important for their activity and stability.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Annual Research Report
  • Research Products

    (14 results)

All 2018 2017 2016

All Journal Article (4 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 4 results,  Acknowledgement Compliant: 1 results,  Open Access: 2 results) Presentation (10 results) (of which Invited: 1 results)

  • [Journal Article] Heterologous expression, purification and characterization of human β-1,2-N-acetylglucosaminyltransferase II using a silkworm-BmNPV bacmid system.2018

    • Author(s)
      Takatsugu Miyazaki, Tatsuya Kato, and Enoch Y. Park
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 印刷中 Issue: 1 Pages: 15-22

    • DOI

      10.1016/j.jbiosc.2018.01.011

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Paenibacillus sp. 598K 6-α-glucosyltransferase is essential for cycloisomaltooligosaccharide synthesis from α-(1→4)-glucan2017

    • Author(s)
      Hitomi Ichinose, Ryuichiro Suzuki, Takatsugu Miyazaki, Keitarou Kimura, Mitsuru Momma, Nobuhiro Suzuki, Zui Fujimoto, Atsuo Kimura, Kazumi Funane
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 101 Issue: 10 Pages: 4115-4128

    • DOI

      10.1007/s00253-017-8174-z

    • NAID

      120006771683

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] N-Glycan Modification of a Recombinant Protein via Coexpression of Human Glycosyltransferases in Silkworm Pupae2017

    • Author(s)
      Tatsuya Kato, Natsumi Kako, Kotaro Kikuta, Takatsugu Miyazaki, Sachiko Kondo, Hirokazu Yagi, Koichi Kato and Enoch Y. Park
    • Journal Title

      Sci. Rep.

      Volume: 7 Issue: 1 Pages: 1409-1409

    • DOI

      10.1038/s41598-017-01630-6

    • NAID

      120006228195

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Insulin-like peptide 3 expressed in the silkworm possesses intrinsic disulfide bonds and full biological activity2017

    • Author(s)
      Miyazaki Takatsugu、Ishizaki Masaaki、Dohra Hideo、Park Sungjo、Terzic Andre、Kato Tatsuya、Kohsaka Tetsuya、Park Enoch Y.
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 17339-17339

    • DOI

      10.1038/s41598-017-17707-1

    • NAID

      120006371042

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 哺乳類内在性色素結合型シアノバクテリオクロムの分子機構2018

    • Author(s)
      伏見 圭司, 宮崎 剛亜, 山本 達郎, 竹田 百花, 桑崎 勇人, 中嶋 隆浩, 榎本 元, 池内 昌彦, 朴 龍洙, 佐藤 守俊, 成川 礼
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] カイコーバクミド遺伝子発現系を用いた熱帯熱マラリア原虫に対するワクチン開発の基盤構築2018

    • Author(s)
      稲垣 裕, 林谷 美貴子, 宮崎 剛亜, デオ ヴィピン クマル, Murhandarwati Elsa Heridiana, 加藤 竜也, 朴 龍洙
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] N結合型糖鎖プロセシング酵素と相同性を有する細菌由来酵素の構造と機能2018

    • Author(s)
      宮崎 剛亜
    • Organizer
      第19回 静岡ライフサイエンスシンポジウム
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] ヒト由来糖転移酵素の共発現によるカイコ発現エリスロポエチンのN型糖鎖構造改変2017

    • Author(s)
      堀場 早紀, 加子 夏未, 宮崎 剛亜, 加藤 竜也, 近藤 幸子, 加藤 晃一, 矢木 宏和, 朴 龍洙
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都女子大学(京都府京都市)
    • Year and Date
      2017-03-18
    • Related Report
      2016 Annual Research Report
  • [Presentation] カイコ‐BmNPVバクミド発現系を用いたN-アセチルグルコサミン転移酵素IIの発現と性質の解析2017

    • Author(s)
      宮崎 剛亜, 加藤 竜也, 朴 龍洙
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] カイコで発現させたヒトエリスロポエチンの糖転移酵素共発現によるN型糖鎖構造改変2017

    • Author(s)
      堀場 早紀, 加子 夏未, 宮崎 剛亜, 加藤 竜也, 近藤 幸子, 矢木 宏和, 加藤 晃一, 朴 龍洙
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] カイコ由来N-acetylgalactosaminyltransferaseの基質特異性2017

    • Author(s)
      宮下 龍之介, 宮崎 剛亜, 加藤 竜也, 朴 龍洙
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] カイコ‐BmNPVバクミド発現系を用いた熱帯熱マラリア原虫抗原提示ウイルス様粒子の作製2017

    • Author(s)
      稲垣 裕, 朴 龍洙, 加藤 竜也, 宮崎 剛亜, Deo Vipin Kumar
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 哺乳類内在色素結合型シアノバクテリオクロムを基盤とした汎用的光遺伝学ツールの開発2017

    • Author(s)
      伏見 圭司, 竹田 百花, 山本 達郎, 宮崎 剛亜, 桑崎 勇人, 中嶋 隆浩, 榎本 元, 池内 昌彦, 朴 龍洙, 佐藤 守俊, 成川 礼
    • Organizer
      藍藻の分子生物学2017
    • Related Report
      2017 Annual Research Report
  • [Presentation] カイコ由来糖転移酵素 GnTII 及び GalT オルソログの機能解析2016

    • Author(s)
      宮下 龍之介, 宮崎 剛亜, 加藤 竜也, 朴 龍洙
    • Organizer
      第68回日本生物工学会大会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2016-09-30
    • Related Report
      2016 Annual Research Report

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Published: 2016-09-02   Modified: 2019-03-29  

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