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Development of photochemical cloning for DNA-binding proteins

Research Project

Project/Area Number 23510286
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Chemical biology
Research InstitutionSuzuka University of Medical Science

Principal Investigator

SADAKANE Yutaka  鈴鹿医療科学大学, 薬学部, 教授 (60293304)

Co-Investigator(Kenkyū-buntansha) OHTSUKA Isao  九州保健福祉大学, 薬学部, 准教授 (20389589)
Project Period (FY) 2011 – 2013
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords光反応 / 分析化学 / コンフォメーション病 / DNA結合タンパク質 / ゲルシフトアッセイ / プロテオーム / バイオテクノロジー / 質量分析 / プロテーム
Research Abstract

We developed the new method for analysis of DNA-binding proteins, which play the various physiological roles such as transcriptional regulation, using photo-reactive compound, which is able to form the covalent bond by light irradiation. Our photochemical gel-shift assay reveals that octamer DNA binds several proteins of nuclear extract. We further examined the photochemical cloning methods combining with molecular biological technique, phage panning or instrumental analysis of mass spectrometry. We also analyzed expression pattern and fibrillation of cranial nerve disease, which is a target disease for photochemical methods in this study.

Report

(4 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • 2011 Research-status Report
  • Research Products

    (33 results)

All 2014 2013 2012 2011 Other

All Journal Article (8 results) (of which Peer Reviewed: 3 results) Presentation (23 results) (of which Invited: 1 results) Remarks (2 results)

  • [Journal Article] Disruption of zinc homeostasis and the pathogenesis of senile dementia2014

    • Author(s)
      M. Kawahara, D. Mizuno, H. Koyama, K. Konoha, S. Ohkawara,, Y. Sadakane
    • Journal Title

      Metallomics

      Volume: 6 Pages: 209-219

    • Related Report
      2013 Final Research Report
  • [Journal Article] Disruption of zinc homeostasis and the pathogenesis of senile dementia2014

    • Author(s)
      M. Kawahara, D. Mizuno, H. Koyama, K. Konoha, S. Ohkawara, Y. Sadakane
    • Journal Title

      Metallomics

      Volume: 6 Issue: 2 Pages: 209-216

    • DOI

      10.1039/c3mt00257h

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] D-Histidine and L-histidine attenuate zinc-induced neuronal death in GT1-7 cells2013

    • Author(s)
      M. Kawahara, Y. Sadakane, H. Koyama, K. Konoha, S. Ohkawara
    • Journal Title

      Metallomics

      Volume: 5 Issue: 5 Pages: 453-460

    • DOI

      10.1039/c3mt20264j

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Experimental conditions for analyzing carbohydrate-binding protein using photoreactive lactose ligand prepared with AffiLight CHO2012

    • Author(s)
      Y. Sadakane, M. Yamazi, I. Otsuka
    • Journal Title

      Photomedicine and Photobiology

      Volume: 34 Pages: 37-38

    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Journal Article] Determination of rate constants for beta-linkage isomerization of three specific aspartyl residues in recombinant human alpha A-crystallin protein by reversed-phase HPLC2011

    • Author(s)
      Y. Sadakane, N. Fujii, K. Nakagomi
    • Journal Title

      J Chromatogr. B.

      Volume: 879 Pages: 3240-3246

    • Related Report
      2013 Final Research Report 2011 Research-status Report
  • [Journal Article] Effects of trace metals on the conformational changes of amyloidogenic proteins and their neurotoxicity2011

    • Author(s)
      M. Kawahara, H. Koyama, Y. Sadakane
    • Journal Title

      Biomed. Res.Trace Elements

      Volume: 22 Pages: 7-14

    • NAID

      10028117958

    • Related Report
      2013 Final Research Report
  • [Journal Article] The effect of the structural differences in the reducing terminus of sugars on the binding affinity of carbohydrates and proteins analyzed using photoaffinity labeling2011

    • Author(s)
      I. Ohtsuka, Y. Sadakane, M. Higuchi, N. Hada, J. Hada, N. Kakiuchi, A. Sakushima
    • Journal Title

      Bioorg Med. Chem.

      Volume: 19 Pages: 894-899

    • Related Report
      2013 Final Research Report
  • [Journal Article] The effect of the structural differences in the reducing terminus of sugars on the binding affinity of carbohydrates and proteins analyzed using photoaffinity labeling.2011

    • Author(s)
      Isao Ohtsuka, Yutaka Sadakane, Mari Higuchi, Noriyasu Hada, Junko Hada, Nobuko Kakiuchi, Akiyo Sakushima
    • Journal Title

      Bioorg Med. Chem.

      Volume: 19 Issue: 2 Pages: 894-899

    • DOI

      10.1016/j.bmc.2010.11.067

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Presentation] 光反応性糖鎖ミメティックペプチドの合成とレクチン解析への応用2014

    • Author(s)
      定金豊, 松下秀峰, 山路充宏, 大塚功
    • Organizer
      日本薬学会第134年会
    • Place of Presentation
      熊本
    • Related Report
      2013 Annual Research Report 2013 Final Research Report
  • [Presentation] 脳血管性認知症予防・治療薬としてのカルノシンの作用メカニズム解明2014

    • Author(s)
      水野 大,高橋 千紘,金子 奈央,定金 豊,川原 正博
    • Organizer
      日本薬学会第134年会
    • Place of Presentation
      熊本
    • Related Report
      2013 Annual Research Report
  • [Presentation] プリオンタンパク質ペプチド断片のN末アミノ酸の構造変化と線維化速度への影響2013

    • Author(s)
      宇佐美亜由子, 山本理香子, 定金豊
    • Organizer
      第59回日本薬学会東海支部大会
    • Place of Presentation
      名古屋
    • Year and Date
      2013-07-06
    • Related Report
      2013 Final Research Report
  • [Presentation] プリオンタンパク質ペプチド断片のN末構造変化と線維化速度への影響2013

    • Author(s)
      定金豊, 宇佐美亜由子, 山本理香子
    • Organizer
      日本薬学会第133年会
    • Place of Presentation
      横浜
    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Presentation] プリオンタンパク質ペプチド断片のN末アミノ酸の構造変化と線維化速度への影響2013

    • Author(s)
      宇佐美亜由子,山本理香子,定金 豊
    • Organizer
      第59回日本薬学会東海支部大会
    • Place of Presentation
      名古屋
    • Related Report
      2013 Annual Research Report
  • [Presentation] 光反応性糖鎖リガンドを用いた結合解析法の効率化2012

    • Author(s)
      定金豊, 山路充宏, 大塚功
    • Organizer
      日本病院薬剤師会東海ブロック・日本薬学会東海支部 合同学術大会2012
    • Place of Presentation
      岐阜
    • Year and Date
      2012-11-18
    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Presentation] 脳疾患に関連するペプチド中のアスパラギンおよびアスパラギン酸残基の構造変化と生理活性2012

    • Author(s)
      定金豊, 小山裕也, 川原正博
    • Organizer
      第57回日本薬学会東海支部大会
    • Place of Presentation
      名古屋
    • Year and Date
      2012-07-09
    • Related Report
      2013 Final Research Report 2011 Research-status Report
  • [Presentation] アルツハイマーβペプチド断片のN末端の構造変化による線維化速度への影響2012

    • Author(s)
      定金豊, 宇佐美亜由子, 山本理香子
    • Organizer
      第58回日本薬学会東海支部大会
    • Place of Presentation
      静岡
    • Year and Date
      2012-07-07
    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Presentation] プリオンタンパク質断片ペプチド(106-126)のN末およびアスパラギン残基の構造変化によるアミロイド線維化速度への影響2012

    • Author(s)
      定金豊, 宇佐美亜由子, 山本理香子
    • Organizer
      第8回Dアミノ酸研究会
    • Place of Presentation
      滋賀
    • Related Report
      2013 Final Research Report
  • [Presentation] 還元末端が閉環した構造をもつ光反応性糖鎖リガンドの合成と結合解析法の確立2012

    • Author(s)
      定金豊, 大塚功, 山路充宏, 畑中保丸
    • Organizer
      第34回日本光医学・光生物学会
    • Place of Presentation
      神戸
    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Presentation] Synthesis of new molecular tools for elucidation of carbohydrates roles by photoaffinity labeling: Carbohydrate - Carbohydrate-protein interactions are affected by different of glycosidic formations2012

    • Author(s)
      I. Ohtsuka, Y. Sadakane, N. Hada, M. Higuchi, N. Kakiuchi
    • Organizer
      International Carbohydrate Symposium
    • Place of Presentation
      Spain
    • Related Report
      2013 Final Research Report
  • [Presentation] Molecular mechanism of zinc-induced neurodegeneration: involvement of calcium dyshomeostasis and carnosine2012

    • Author(s)
      S. OHKAWARA, H. KOYAMA, Y. SADAKANE, M. KAWAHARA
    • Organizer
      第22回金属の関与する生体関連反応シンポジウム
    • Place of Presentation
      金沢
    • Related Report
      2013 Final Research Report
  • [Presentation] アルツハイマーβペプチドのN末側アミノ酸の構造的変化と線維化への影響2012

    • Author(s)
      定金豊, 小山裕也, 川原正博
    • Organizer
      日本薬学会第132年会
    • Place of Presentation
      札幌
    • Related Report
      2013 Final Research Report
  • [Presentation] PrP106-126の高次構造変化に重要な配列2012

    • Author(s)
      小山裕也, 坂口哲陛, 定金豊, 川原正博
    • Organizer
      日本薬学会第132年会
    • Place of Presentation
      札幌
    • Related Report
      2013 Final Research Report
  • [Presentation] 光アフィニティーラベル法による新規分子ツールの開発~糖鎖結合様式が認識に及ぼす影響を検証~2012

    • Author(s)
      大塚功, 定金豊, 羽田紀康, 羽田純子, 新垣達司, 樋口真里, 垣内信子
    • Organizer
      日本薬学会第132年会
    • Place of Presentation
      札幌
    • Related Report
      2013 Final Research Report 2011 Research-status Report
  • [Presentation] Synthesis of new molecular tools for elucidation of carbohydrates roles by photoaffinity labeling: Carbohydrate – Carbohydrate-protein interactions are affected by different of glycosidic formations.2012

    • Author(s)
      I. Ohtsuka; Y. Sadakane; N. Hada, M. Higuchi; N. Kakiuchi
    • Organizer
      International Carbohydrate Symposium
    • Place of Presentation
      Spain
    • Related Report
      2012 Research-status Report
  • [Presentation] プリオンタンパク質断片ペプチド(106-126)のN末およびアスパラギン残基の構造変化によるアミロイド線維化速度への影響2012

    • Author(s)
      定金 豊、宇佐美亜由子、山本理香子
    • Organizer
      第8回Dアミノ酸研究会
    • Place of Presentation
      滋賀
    • Related Report
      2012 Research-status Report
  • [Presentation] PrP106-126 の高次構造変化に重要な配列2012

    • Author(s)
      小山 裕也,坂口 哲陛,定金 豊,川原 正博
    • Organizer
      日本薬学会第132年会
    • Place of Presentation
      札幌
    • Related Report
      2011 Research-status Report
  • [Presentation] アルツハイマー β ペプチドの N 末側アミノ酸の構造的変化と線維化への影響2012

    • Author(s)
      定金 豊,小山 裕也,川原 正博
    • Organizer
      日本薬学会第132年会
    • Place of Presentation
      札幌
    • Related Report
      2011 Research-status Report
  • [Presentation] 神経疾患に関わるペプチド断片の機能とアミノ酸レベルの構造変化:アミロイド線維化と細胞毒性への影響2011

    • Author(s)
      定金豊
    • Organizer
      京都大学原子炉実験所・専門研究会「第4回タンパク質の異常凝集とその防御・修復機構に関する研究会」
    • Place of Presentation
      大阪
    • Year and Date
      2011-11-11
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] 部位特異的カルベン発生型DNA, ペプチド, タンパク質の作製ツールの開発2011

    • Author(s)
      定金豊, 畑中保丸
    • Organizer
      第33回日本光医学・光生物学会
    • Place of Presentation
      大阪
    • Year and Date
      2011-07-23
    • Related Report
      2013 Final Research Report
  • [Presentation] 神経疾患に関わるペプチド断片の機能とアミノ酸レベルの構造変化:アミロイド線維化と細胞毒性への影響2011

    • Author(s)
      定金 豊
    • Organizer
      京都大学原子炉実験所・専門研究会 「第4回 タンパク質の異常凝集とその防御・修復機構に関する研究会」(招待講演)
    • Place of Presentation
      大阪
    • Related Report
      2011 Research-status Report
  • [Presentation] 部位特異的カルベン発生型DNA,ペプチド,タンパク質の作製ツールの開発2011

    • Author(s)
      定金 豊,畑中保丸
    • Organizer
      第33回 日本光医学・光生物学会
    • Place of Presentation
      大阪
    • Related Report
      2011 Research-status Report
  • [Remarks]

    • URL

      http://www.d4.dion.ne.jp/~sadafami/

    • Related Report
      2013 Final Research Report
  • [Remarks]

    • URL

      http://www.d4.dion.ne.jp/~sadafami/

    • Related Report
      2011 Research-status Report

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Published: 2011-08-05   Modified: 2019-07-29  

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