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Development of novel analytical methods for phosphoproteomics by using a phosphate-binding tag molecule.

Research Project

Project/Area Number 19390011
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionHiroshima University

Principal Investigator

KOIKE Tohru  Hiroshima University, 大学院・医歯薬学総合研究科, 教授 (90186586)

Project Period (FY) 2007 – 2009
Project Status Completed (Fiscal Year 2009)
Budget Amount *help
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2009: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2008: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2007: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000)
Keywordsリン酸化プロテオミクス / ケミカルバイオロジー / リン酸化蛋白質 / キナーゼ / フォスファターゼ / プロテオミクス / リン酸化タンパク質 / ナノバイオ / シグナル伝達 / FRET
Research Abstract

By utilizing the Phos-tag molecule and its derivatives, we have developed convenient and reliable methods for the detection and separation of phosphorylated peptides and proteins. In this program, we applied the Phos-tag technology to (1) FRET system for kinase profiling using Phos-tag, (2) phosphate affinity electrophoresis, (3) phosphate affinity column system. We believe that our Phos-tag technology will result in great progress in phosphoproteomics.

Report

(4 results)
  • 2009 Annual Research Report   Final Research Report ( PDF )
  • 2008 Annual Research Report
  • 2007 Annual Research Report
  • Research Products

    (41 results)

All 2009 2008 2007 Other

All Journal Article (19 results) (of which Peer Reviewed: 19 results) Presentation (13 results) Book (5 results) Remarks (4 results)

  • [Journal Article] Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysates2009

    • Author(s)
      E. Kinoshita-Kikuta, E. Kinoshita, T. Koike
    • Journal Title

      Analytical Biochemistry 389

      Pages: 83-85

    • NAID

      120002023644

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phospho-peptides2009

    • Author(s)
      K. Takiyama, E. Kinoshita, E. Kinoshita-Kikuta, Y. Fujioka, Y. Kubo, T. Koike
    • Journal Title

      Analytical Biochemistry 388

      Pages: 235-241

    • NAID

      120002023643

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] Two-dimensional phosphate affinity gel electrophoresis for the analysis of phosphoprotein isotypes2009

    • Author(s)
      E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, Y. Aoki, S. Ohie, Y. Mouri, T. Koike
    • Journal Title

      Electrophoresis 30

      Pages: 550-559

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE2009

    • Author(s)
      E. Kinoshita, E. Kinoshita-Kikuta, T. Koike
    • Journal Title

      Nature Protocols 4

      Pages: 1513-1521

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agarose2009

    • Author(s)
      Eiji Kinoshita
    • Journal Title

      Proteomics 9

      Pages: 4098-4101

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptides2009

    • Author(s)
      Kei Takiyama
    • Journal Title

      Analytical Biochemistry 388

      Pages: 235-241

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysates2009

    • Author(s)
      Eiji Kinoshita
    • Journal Title

      Analytical Biochemistry 389

      Pages: 83-85

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Two-dimensional Phosphate Affinity Gel Electrophoresis for the Analysis of Phosphoprotein Isotypes2009

    • Author(s)
      Eiji Kinoshita
    • Journal Title

      Electrophoresis 30

      Pages: 550-559

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A mobility-shift detection method for DNA methylation analysis using phosphate-affinity polyacrylamide gel electrophoresis2008

    • Author(s)
      E. Kinoshita-Kikuta, E. Kinoshita, T. Koike
    • Journal Title

      Analytical Biochemistry 378

      Pages: 102-104

    • NAID

      120001092444

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate-affinity SDS-PAGE2008

    • Author(s)
      E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, S. Yamada, H. Nakamura, Y. Shiro, Y. Aoki, K. Okita, T. Koike
    • Journal Title

      Proteomics 8

      Pages: 2994-3003

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] FANCI phosphorylation functions as a molecular switch to turn on the Fanconi anemia pathway2008

    • Author(s)
      M. Ishiai, H. Kitao, A. Smogorzewska, J, Tomida, A. Kinomura, E. Uchida, A. Saberi, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, S. Tashiro, S.J. Elledge, M. Takata
    • Journal Title

      Nature Structural & Molecular Biology 15

      Pages: 1138-1146

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] A mobility-shift detection method for DNA methylation analysisusing phosphate-affinity polyacrylamide gel electrophoresis2008

    • Author(s)
      Emiko Kinoshita-Kikuta
    • Journal Title

      Analytical Biochemistry 378

      Pages: 102-104

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate-affinity SDS-PAGE2008

    • Author(s)
      Eiji Kinoshita
    • Journal Title

      Proteomics 8

      Pages: 2994-3003

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Separation of a phosphorylated-His protein using phosphate-affinity polyacrylamide gel2007

    • Author(s)
      S. Yamada, H. Nalkamura, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, Y. Shiro
    • Journal Title

      Analytical Biochemistry 360

      Pages: 160-162

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] A single nucleotide polymorphism genotyping method using phosphate-affinity polyacrylamide gel electrophoresis Analytical2007

    • Author(s)
      E. Kinoshita, E. Kinoshita-Kikuta, T. Koike
    • Journal Title

      Biochemistry 361

      Pages: 294-298

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] Label-free kinase profiling using phosphate-affinity polyacrylamide gel electrophoresis2007

    • Author(s)
      E. Kinoshita-Kikuta, Y. Aoki, E. Kinoshita, T. Koike
    • Journal Title

      Molecular & Cellular Proteomics 6

      Pages: 356-366

    • NAID

      120000878140

    • Related Report
      2009 Final Research Report
    • Peer Reviewed
  • [Journal Article] Identification on Membrane and Characterization of Phosphoproteins Using an Alkoxide-bridged Metal Complex as a Phosphate-binding Tag Molecule2007

    • Author(s)
      Tsuyoshi Nakanishi
    • Journal Title

      Journal of Biomolecular Techniques 18

      Pages: 278-286

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] キナーゼプロファイリングのための新しいリン酸アフィニティー電気泳動法2007

    • Author(s)
      木下英司
    • Journal Title

      生物物理化学 51

      Pages: 199-206

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A single nucleotide polymorphism genotyping method using phosphate-affinity polyacrylamide gel electrophoresis2007

    • Author(s)
      Eiji Kinoshita
    • Journal Title

      Analytical Biochemistry 361

      Pages: 294-298

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] 改良型リン酸基親和性電気泳動法による200kDa以上の高分子量タンパク質のリン酸化解析2009

    • Author(s)
      木下恵美子
    • Organizer
      第82回日本生化学会大会
    • Place of Presentation
      兵庫県神戸市
    • Year and Date
      2009-10-22
    • Related Report
      2009 Annual Research Report
  • [Presentation] リン酸基親和性電気泳動法を用いたヒスチジン・アスパラギン酸リン酸化タンパク質の定量解析法2009

    • Author(s)
      柳原志穂, 木下恵美子, 木下英司, 中神明, 小池透
    • Organizer
      第82回日本生化学会大会合同大会
    • Place of Presentation
      神戸
    • Year and Date
      2009-10-21
    • Related Report
      2009 Final Research Report
  • [Presentation] 蛋白質リン酸化解析のためのリン酸基親和性2次元電気泳動法2009

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第60回日本電気泳動学会総会
    • Place of Presentation
      長野
    • Year and Date
      2009-09-18
    • Related Report
      2009 Final Research Report
  • [Presentation] Phos-tag固定化ポリマーを利用したシグナル伝達解析における選択的蛋白質リン酸化検出のための新技術2009

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      日本ヒトプロテオーム機構第7回大会
    • Place of Presentation
      東京
    • Year and Date
      2009-07-27
    • Related Report
      2009 Final Research Report
  • [Presentation] A novel FRET system for analysis of peptide phosphorylation status2008

    • Author(s)
      Kei Takiyama
    • Organizer
      第81回日本生化学会大会
    • Place of Presentation
      兵庫県神戸市
    • Year and Date
      2008-12-10
    • Related Report
      2008 Annual Research Report
  • [Presentation] フォスタグ結合型新規リン酸基アフィニティービーズを利用した細胞抽出液からのリン酸化蛋白質の分離と精製2008

    • Author(s)
      木下英司, 木下恵美子, 山田篤志, 井上知香, 小池透
    • Organizer
      日本ヒトプロテオーム機構第6回大会
    • Place of Presentation
      大阪
    • Year and Date
      2008-07-29
    • Related Report
      2009 Final Research Report
  • [Presentation] ウエスタンブロッティング膜に転写されたリン酸化蛋白質の検出とダイレクト膜上質量分析2008

    • Author(s)
      木下英司, 木下恵美子, 中西豪, 安藤英治, 古田大, 綱澤進, 西村紀, 小池透
    • Organizer
      日本薬学会第128年会
    • Place of Presentation
      横浜
    • Year and Date
      2008-03-26
    • Related Report
      2009 Final Research Report
  • [Presentation] Phosphate affinity 2-D gel electro- phoresis for the analysis of phospho- proteins2007

    • Author(s)
      E. Kinoshita, E. Kinoshita- Kikuta, Y. Aoki, S. Ohie, Y. Mouri, Mamoru Matsubara, T. Koike
    • Organizer
      第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
    • Place of Presentation
      横浜
    • Year and Date
      2007-12-12
    • Related Report
      2009 Final Research Report
  • [Presentation] Phosphate affinity gel electrophoresis using a phosphate-binding tag molecule, Phos-tag, for protein kinase and phosphatase profiling2007

    • Author(s)
      木下英司
    • Organizer
      第4回武田科学振興財団薬科学シンポジウム
    • Place of Presentation
      東京都港区
    • Year and Date
      2007-12-03
    • Related Report
      2007 Annual Research Report
  • [Presentation] 遺伝子変異を認識する機能性分子を用いた遺伝子診断技術の開発2007

    • Author(s)
      木下英司, 木下恵美子, 由元実希, 小池透
    • Organizer
      第58回日本電気泳動学会総会
    • Place of Presentation
      宇部
    • Year and Date
      2007-11-07
    • Related Report
      2009 Final Research Report
  • [Presentation] リン酸化プロテオーム研究の新戦略2007

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      日本ヒトプロテオーム機構第5回大会
    • Place of Presentation
      東京
    • Year and Date
      2007-07-30
    • Related Report
      2009 Final Research Report
  • [Presentation] リン酸基結合ナノ分子を利用した新しい二次元電気泳動法の開発2007

    • Author(s)
      木下英司, 木下恵美子, 大家志織, 松原守, 小池透
    • Organizer
      第49回生化学会中四国支部例会
    • Place of Presentation
      高知
    • Year and Date
      2007-05-19
    • Related Report
      2009 Final Research Report
  • [Presentation] 消化プロセスで生じる創傷治癒ホルモン・リゾホスファチジン酸による消化管組織修復2007

    • Author(s)
      中保, 堀内剛, 平野薫, 徳村彰, 盛重純一, 小池透, Mandi Murph, Gordon Mills
    • Organizer
      里内清, 第49回生化学会中四国支部例会
    • Place of Presentation
      高知
    • Year and Date
      2007-05-19
    • Related Report
      2009 Final Research Report
  • [Book] 細胞内リン酸化タンパク質を高精度に検出するためのPhos-tagビーズによるサンプル前処理法,細胞工学2009

    • Author(s)
      木下英司,木下恵美子,小池透
    • Total Pages
      938
    • Publisher
      秀潤社
    • Related Report
      2009 Final Research Report
  • [Book] Phosphate-affinity Gel Electrophoresis Using a Phos-tag Molecule for Phosphoproteome Study2009

    • Author(s)
      E.Kinoshita, E.Kinoshita-Kikuta, T.Koike
    • Total Pages
      104
    • Publisher
      Current Proteomics
    • Related Report
      2009 Final Research Report
  • [Book] リン酸基アフィニティー電気泳動法を利用した遺伝子診断法,実験医学2007

    • Author(s)
      木下英司,木下恵美子,小池透
    • Total Pages
      3033
    • Publisher
      羊土社
    • Related Report
      2009 Final Research Report
  • [Book] 金属イオンの特性を利用したリン酸化タンパク質の特異的認識と検出2007

    • Author(s)
      木下英司,木下恵美子,小池透
    • Total Pages
      1897
    • Publisher
      薬学雑誌
    • Related Report
      2009 Final Research Report
  • [Book] リン酸基親和性クロマトグラフィーを用いた細胞内のリン酸化タンパク質の網羅的精製,バイオテクノロジージャーナル2007

    • Author(s)
      木下英司,木下恵美子,小池透
    • Total Pages
      217
    • Publisher
      羊土社
    • Related Report
      2009 Final Research Report
  • [Remarks]

    • URL

      http://home.hiroshima-u.ac.jp/tkoike/

    • Related Report
      2009 Final Research Report
  • [Remarks]

    • URL

      http://home.hiroshima-u.ac.jp/tkoike/

    • Related Report
      2009 Annual Research Report
  • [Remarks]

    • URL

      http://home.hiroshima-u.ac.jp/tkoike/

    • Related Report
      2008 Annual Research Report
  • [Remarks]

    • URL

      http://home.hiroshima-u.ac.jp/tkoike/

    • Related Report
      2007 Annual Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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