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Development of anti-Trypanosoma drug based on structural and functional analysis of novel L-amino acid dehydrogenase

Research Project

Project/Area Number 24780105
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Applied biochemistry
Research InstitutionTokai University

Principal Investigator

YONEDA Kazunari  東海大学, 農学部, 講師 (00469397)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords酵素化学 / L-スレオニン酸脱水素酵素 / NAD / トリパノソーマ / クローニング / 結晶化 / 脱水素酵素 / L-スレオニン
Research Abstract

L-Threonine dehydrogenase (ThrDH) gene from the pathogenic parasite Trypanosoma brucei was amplified using the overlapping primers, and ligated with the expression vector pCold TF. Escherichia coli was then transformed with the vector and induced by adding 1.0 mM IPTG to the medium. The gene was overexpressed in E. coli, and its product was purified and characterized. SDS-PAGE showed the subunit molecular mass of T. brucei ThrDH to be about 81 kDa, which was consistent with the molecular weight calculated from the amino acid sequence. The slightly NAD-dependent ThrDH activity was determined.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (24 results)

All 2013 2012 Other

All Journal Article (10 results) (of which Peer Reviewed: 10 results) Presentation (12 results) (of which Invited: 2 results) Remarks (2 results)

  • [Journal Article] Crystallization and preliminary X-ray analysis of L-serine 3-dehydrogenase complexed with NADP^+ from the hyperthermophilic archaeon Pyrobaculum calidifontis2013

    • Author(s)
      Yoneda K, Sakuraba H, Araki T, Shibata T, Nikki T, Ohshima T
    • Journal Title

      Acta Crystallographica Section F

      Volume: Vol. 69 Pages: 134-136

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and preliminary X-ray analysis of L-serine 3-dehydrogenase complexed with NADP+ from the hyperthermophilic archaeon Pyrobaculum calidifontis,2013

    • Author(s)
      Yoneda, K., Sakuraba, H., Araki, T., Shibata, T., Nikki, T. & Ohshima, T.
    • Journal Title

      Acta Cryst.

      Volume: F69 Issue: 2 Pages: 134-136

    • DOI

      10.1107/s1744309112051391

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Crystal Structure of the binary and ternary complexes of an archaeal UDP-galactose 4-epimerase-like L-threonine dehydrogenase from Thermoplasma volcanium2012

    • Author(s)
      Yoneda K, Sakuraba H, Araki T, Ohshima T
    • Journal Title

      J. Biol. Chem.

      Volume: Vol. 287 Pages: 12966-12974

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Expression, purification, crystallization and preliminary X-ray analysis of NAD(P)H-dependent carbonyl reductase specifically expressed in the thyroidectomized chicken fatty liver2012

    • Author(s)
      Yoneda K, Fukuda Y, Shibata T, Araki T, Nikki T, Sakuraba H, Ohshima T
    • Journal Title

      Acta Crystallographica Section F

      Volume: Vol. 68 Pages: 1568-1570

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystal Structure of Novel Dye-linked L-Proline Dehydrogenase from Hyperthermophilic Archaeon Aeropyrum pernix2012

    • Author(s)
      Sakuraba H, Satomura T, Kawakami R, Kim K, Hara Y, Yoneda K, Ohshima T
    • Journal Title

      J. Biol. Chem.

      Volume: Vol. 287 Pages: 20070-20080

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structure of a UDP-glucose dehydrogenase from the hyperthermophilic archaeon Pyrobaculum islandicum2012

    • Author(s)
      Sakuraba H, Kawai T, Yoneda K, Ohshima T
    • Journal Title

      Acta Crystallographica Section

      Volume: Vol. 68 Pages: 1003-1007

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystal Structure of Binary and Ternary Complexes of Archaeal UDP-galactose 4-Epimerase-like L-Threonine Dehydrogenase from Thermoplasma volcanium.2012

    • Author(s)
      K.Yoneda
    • Journal Title

      The Journal of Biological Chemistry

      Volume: VOL. 287 Issue: 16 Pages: 12966-12974

    • DOI

      10.1074/jbc.m111.336958

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Expression, purification, crystallization and preliminary X-ray analysis of NAD(P)H-dependent carbonyl reductase specifically expressed in thyroidectomized chicken fatty liver2012

    • Author(s)
      Kazunari Yoneda, Yudai Fukuda, Takeshi Shibata, Tomohiro Araki, Takahiro Nikki, Haruhiko Sakurba, Toshihisa Ohshima
    • Journal Title

      Acta Crystallographica

      Volume: F68 Issue: 12 Pages: 1568-1570

    • DOI

      10.1107/s1744309112046453

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Crystal Structure of Novel Dye-linked L-Proline Dehydrogenase from Hyperthermophilic Archaeon Aeropyrum pernix2012

    • Author(s)
      Haruhiko Sakuraba, Takenori Satomura, Ryushi Kawakami, Kwang Kim, Yusuke Hara, Kazunari Yoneda, Toshihisa Ohshima
    • Journal Title

      Journal of Biological Chemistry

      Volume: 287 Issue: 24 Pages: 20070-20080

    • DOI

      10.1074/jbc.m111.319038

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Structure of a UDP-glucose dehydrogenase from the hyperthermophilic archaeon Pyrobaculum islandicum2012

    • Author(s)
      Sakuraba H., Kawai T., Yoneda K., Ohshima T.
    • Journal Title

      Acta Cryst

      Volume: F68 Issue: 9 Pages: 1003-1007

    • DOI

      10.1107/s1744309112030667

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Presentation] トリパノソーマ由来NAD依存性L-スレオニン脱水素酵素の性質と構造2013

    • Author(s)
      米田一成、櫻庭春彦、荒木朊洋、大島敏久
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館
    • Year and Date
      2013-06-12
    • Related Report
      2013 Final Research Report
  • [Presentation] トリパノソーマ由来NAD依存性L-スレオニン脱水素酵素の機能解析2013

    • Author(s)
      米田一成、櫻庭春彦、荒木朊洋、大島敏久
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      東北大学
    • Year and Date
      2013-03-25
    • Related Report
      2013 Final Research Report
  • [Presentation] 新規な補酵素NAD(P)依存性L-スレオニン脱水素酵素の機能と構造解析2013

    • Author(s)
      米田一成
    • Organizer
      第1回佐賀大学総合分析実験センターシンポジウム
    • Place of Presentation
      佐賀大学
    • Year and Date
      2013-03-18
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] トリパノソーマ由来NAD依存性L-スレオニン脱水素酵素の性質と構造

    • Author(s)
      米田一成、櫻庭 春彦、荒木 朋洋、大島 敏久
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館(鳥取県)
    • Related Report
      2013 Annual Research Report
  • [Presentation] ニワトリ脂肪肝で特異的に発現するNAD(P)H依存性carbonyl redactaseの性質と構造

    • Author(s)
      曽根孟起、福田雄大, 荒木朋洋, 櫻庭春彦, 大島敏久, 米田 一成
    • Organizer
      日本農芸化学会関西・中四国・西日本支部および日本ビタミン学会近畿・中国四国・九州沖縄地区合同大会
    • Place of Presentation
      県立広島大学(広島県)
    • Related Report
      2013 Annual Research Report
  • [Presentation] ニワトリ脂肪肝で特異的に発現するNAD(P)H依存性carbonyl reductaseの分子特性

    • Author(s)
      福田雄大、荒木朋洋,仁木隆博,櫻庭春彦,大島敏久,芝田猛,米田一成
    • Organizer
      日本農芸化学会関西・中四国・西日本支部および日本ビタミン学会近畿・中国四国・九州沖縄地区合同大会
    • Place of Presentation
      県立広島大学(広島県)
    • Related Report
      2013 Annual Research Report
  • [Presentation] 超好熱ア-キアPyrobaculum calidifontis由来NAD(P)依存性L-セリン 3-脱水素酵素の機能・構造解析

    • Author(s)
      米田一成、櫻庭 春彦、荒木 朋洋、大島 敏久
    • Organizer
      日本農芸化学会2014年度大会
    • Place of Presentation
      明治大学(東京都)
    • Related Report
      2013 Annual Research Report
  • [Presentation] トリパノソーマ由来NAD依存性L-スレオニン脱水素酵素の機能解析

    • Author(s)
      米田一成
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      東北大学(宮城県)
    • Related Report
      2012 Research-status Report
  • [Presentation] ニワトリ脂肪肝で特異的に発現する機能未知タンパク質の機能解析

    • Author(s)
      米田一成
    • Organizer
      日本ビタミン学会第64回大会
    • Place of Presentation
      長良川国際会議場(岐阜県)
    • Related Report
      2012 Research-status Report
  • [Presentation] ニワトリ脂肪肝で特異的に発現するNAD(P)H依存性carbonyl reductaseの機能解析

    • Author(s)
      福田雄大
    • Organizer
      平成24年度日本農芸化学会西日本支部および日本栄養・食糧学会九州・沖縄支部合同大会
    • Place of Presentation
      鹿児島大学(鹿児島県)
    • Related Report
      2012 Research-status Report
  • [Presentation] Crystal structure and application of hyperthermophilic lysine 6-dehydrogenase as an NAD-dependent amine dehydrogenase

    • Author(s)
      T. Ohshima
    • Organizer
      12th Japan-China-Korea Joint Symposium on Enzyme Engineering
    • Place of Presentation
      金沢エクセルホテル東急(石川県)
    • Related Report
      2012 Research-status Report
  • [Presentation] 新規な補酵素NAD(P)依存性L-スレオニン脱水素酵素の機能と構造解析

    • Author(s)
      米田一成
    • Organizer
      第1回佐賀大学総合分析実験センターシンポジウム
    • Place of Presentation
      佐賀大学(佐賀県)
    • Related Report
      2012 Research-status Report
    • Invited
  • [Remarks]

    • URL

      http://www2.ktokai-u.ac.jp/~nougaku/Bio/yoneda/yoneda.htm

    • Related Report
      2013 Final Research Report
  • [Remarks] 生物化学研究室HP

    • URL

      http://www2.ktokai-u.ac.jp/~nougaku/Bio/yoneda/yoneda.htm

    • Related Report
      2012 Research-status Report

URL: 

Published: 2013-05-31   Modified: 2019-07-29  

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