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A method for simultaneous detection of dual post-translational modifications using phosphate-affinity electrophoresis

Research Project

Project/Area Number 22790037
Research Category

Grant-in-Aid for Young Scientists (B)

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

Principal Investigator

KINOSHITA Emiko  広島大学, 大学院・医歯薬学総合研究科, 助教 (40379912)

Project Period (FY) 2010 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2010: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywordsリン酸親和性2次元電気泳動法 / Phos-tag SDS-PAGE / 翻訳後修飾 / リン酸化 / ユビキチン化 / βカテニン / 卵白アルブミン / 蛍光ディファレンシャル電気泳動法 / β-カテニン / ナノ材料 / 分子認識 / 蛋白質 / バイオテクノロジー / 分析化学 / フォスタグ / 電気泳動
Research Abstract

The researcher developed a method for simultaneous detection of dual post-translational modifications using two-dimensional electrophoresis based on the original principal of phosphate-affinity electrophoresis, Phos-tag SDS-PAGE. By coupling general SDS-PAGE as the first dimension and Phos-tag SDS-PAGE as the second dimension, dual post-translational modifications of cellular proteins, such as phosphorylation and ubiquitination, were detectable simultaneously on the same gel.

Report

(3 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • Research Products

    (55 results)

All 2012 2011 2010 Other

All Journal Article (25 results) (of which Peer Reviewed: 13 results) Presentation (26 results) Remarks (4 results)

  • [Journal Article] Separation andidentification of four distinctserine-phosphorylation states of ovalbuminby Phos-tag affinity electrophoresis2012

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

      Electrophoresis

      Volume: in press

    • Related Report
      2011 Final Research Report
  • [Journal Article] Highly sensitive detectionof protein phosphorylation by usingimproved Phos-tag Biotin2012

    • Author(s)
      Kinoshita E., Kinoshita-Kikuta E., Sugiyama Y., Fukuda Y., Ozeki T., andKoike T
    • Journal Title

      in press

    • Related Report
      2011 Final Research Report
  • [Journal Article] Phos-tag SDS-PAGEsystems for phosphorylation profiling ofproteins with a wide range of molecularmasses under neutral pH conditions2012

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

      Proteomics

      Volume: 12 Pages: 192-202

    • Related Report
      2011 Final Research Report
  • [Journal Article] 小池透フォスタグケミストリー2012

    • Author(s)
      木下英司, 木下恵美子
    • Journal Title

      生物物理化学

      Volume: 56 Pages: 3-7

    • Related Report
      2011 Final Research Report
  • [Journal Article] 小池透今後の展望1中性条件で泳動を行う改良型Phos-tag SDS-PAGEを用いたリン酸化タンパク質解析2012

    • Author(s)
      木下恵美子, 木下英司
    • Journal Title

      生物物理化学

      Volume: 56 Pages: 41-44

    • Related Report
      2011 Final Research Report
  • [Journal Article] 小池透今後の展望2蛍光性Phos-tagを用いたタンパク質リン酸化反応の解析法2012

    • Author(s)
      木下英司, 木下恵美子, 多幾山敬, 宗村雅男
    • Journal Title

      生物物理化学

      Volume: 56 Pages: 45-49

    • Related Report
      2011 Final Research Report
  • [Journal Article] 小池透プロトコール集フォスタグテクノロジーを用いたリン酸化タンパク質解析法2012

    • Author(s)
      木下恵美子, 木下英司
    • Journal Title

      生物物理化学

      Volume: 56 Pages: 51-75

    • Related Report
      2011 Final Research Report
  • [Journal Article] Phos-tag SDS-PAGE systems for phosphorylation profiling of proteins with a wide range of molecular masses under neutral pH conditions2012

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

      Proteomics

      Volume: 12 Issue: 2 Pages: 192-202

    • DOI

      10.1002/pmic.201100524

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Separation and identification of four distinct serine-phosphorylation states of ovalbumin by Phos-tag affinity electrophoresis2012

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

      Electrophoresis

      Volume: (掲載決定)

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Highly sensitive detection of protein phosphorylation by using improved Phos-tag Biotin2012

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

      Proteomics

      Volume: (掲載決定)

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] フォスタグケミストリー2012

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Journal Title

      生物物理化学

      Volume: 56

    • NAID

      130004809452

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 今後の展望1中性条件で泳動を行う改良型Phos-tag SDS-PAGEを用いたリン酸化タンパク質解析2012

    • Author(s)
      木下恵美子, 木下英司, 小池透
    • Journal Title

      生物物理化学

      Volume: 56

    • NAID

      130004809455

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 今後の展望2蛍光性Phos-tagを用いたタンパク質リン酸化反応の解析法2012

    • Author(s)
      木下英司, 木下恵美子, 多幾山敬, 宗村雅男, 小池透
    • Journal Title

      生物物理化学

      Volume: 56

    • NAID

      130004809456

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] プロトコール集フォスタグテクノロジーを用いたリン酸化タンパク質解析法2012

    • Author(s)
      木下恵美子, 木下英司, 小池透
    • Journal Title

      生物物理化学

      Volume: 56

    • NAID

      130004809457

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Improved Phos-tag SDS-PAG Eunder neutral pH conditions for advancedprotein phosphorylation profiling2011

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

      Proteomics

      Volume: 11 Pages: 319-323

    • Related Report
      2011 Final Research Report
  • [Journal Article] A phos-tag-based fluorescence resonance energy transfer system for theanalysis of the kinase reaction of asubstrate peptide2011

    • Author(s)
      Somura, M., Takiyama, K., Kinoshita-Kikuta E., Kinoshita, E., Koike, T
    • Journal Title

      Analytical Methods

      Volume: 3 Pages: 1303-1309

    • Related Report
      2011 Final Research Report
  • [Journal Article] Zinc(II). cyclen polyacrylamide gel electrophoresis for detection of mutations in short Ade/ Thy-rich DNA fragments2011

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

      AnalyticalBiochemistry

      Volume: 408 Pages: 348-350

    • Related Report
      2011 Final Research Report
  • [Journal Article] A novelphosphate-affinity bead with immobilized Phos-tag for separation and enrichment of phosphopeptides and phosphoproteins2011

    • Author(s)
      Kinoshita-Kikuta, E., Yamada, A., Inoue, C., Kinoshita, E., Koike, T
    • Journal Title

      Journal of Integrated Omics

      Volume: 1 Pages: 157-269

    • Related Report
      2011 Final Research Report
  • [Journal Article] A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the kinase reaction of a substrate peptide2011

    • Author(s)
      M. Somura, K. Takiyama, E. Kinoshita- Kikuta, E. Kinoshita, and T. Koike
    • Journal Title

      Analytical Methods

      Volume: 3 Issue: 6 Pages: 1303-1309

    • DOI

      10.1039/c1ay05016h

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Zinc(II)-cyclen polyacrylamide gel electrophoresis for detection of mutations in short Ade/Thy-rich DNA fragments2011

    • Author(s)
      Kinoshita-Kikuta, E., Eiji Kinoshita, Harada, N., and Koike, T
    • Journal Title

      Anal. Biochem

      Volume: 408 Issue: 2 Pages: 348-350

    • DOI

      10.1016/j.ab.2010.09.003

    • NAID

      120002753732

    • URL

      http://ir.lib.hiroshima-u.ac.jp/00030873

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel phosphate-affinity bead with immobilized Phos-tag for separation and enrichment of phosphopeptides and phosphoproteins2011

    • Author(s)
      Kinoshita-Kikuta, E., Yamada, A., Inoue, C., Eiji Kinoshita, and Koike, T
    • Journal Title

      J. Integrated OMICS

      Volume: 1 Issue: 1 Pages: 157-169

    • DOI

      10.5584/jiomics.v1i1.46

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Zinc(II)-cyclen polyacrylamide gel electrophoresis for detection of mutations in short Ade/Thy-rich DNA fragments.2011

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

      Anal.Biochem.

      Volume: 408 Pages: 348-350

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Improved Phos-tag SDS-PAGE under neutral pH conditions for advancedprotein phosphorylation profiling.2011

    • Author(s)
      Kinoshita, E.
    • Journal Title

      Proteomics

      Volume: 11 Pages: 319-323

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel phosphate-affinity bead with immobilized Phos-tag for separationand enrichment of phosphopeptides and phosphoproteins.2011

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

      J.Integr.Omics

      Volume: 1 Pages: 157-169

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Monitoring method for thealteration of phosphorylation status in ahigh-molecular-mass protein using Phos-tag electrophoresis technology2010

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

      Seikagaku

      Volume: 82 Pages: 857-862

    • Related Report
      2011 Final Research Report
  • [Presentation] Emiko Kinoshita-Kikuta, and Tohru Koike, Phos-tag SDS-PAGEsystems for phosphorylation profiling ofsignaling proteins with a wide range ofmolecular masses under neutral pHconditions2011

    • Author(s)
      Eiji Kinoshita
    • Organizer
      第33回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2011-12-16
    • Related Report
      2011 Final Research Report
  • [Presentation] Phos-tag SDS-PAGE systems for phosphorylation profiling of signaling proteins with a wide range of molecular masses under neutral pH conditions2011

    • Author(s)
      Eiji Kinoshita, Emiko Kinoshita-Kikuta, Tohru Koike
    • Organizer
      第33回日本分子生物学会年会
    • Place of Presentation
      横浜市
    • Year and Date
      2011-12-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] Phos-tagBiotinの開発とその応用2011

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第62回日本電気泳動学会総会
    • Place of Presentation
      横浜市開講記念会館改良型
    • Year and Date
      2011-11-13
    • Related Report
      2011 Final Research Report
  • [Presentation] 改良型Phos-tag Biotinの開発とその応用2011

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第62回日本電気泳動学会総会
    • Place of Presentation
      横浜市
    • Year and Date
      2011-11-13
    • Related Report
      2011 Annual Research Report
  • [Presentation] 亜鉛-Phos-tag SDS-PAGEによる細胞内キナーゼ/フォスファターゼの動態解析2011

    • Author(s)
      木下恵美子
    • Organizer
      第84回日本生化学大会
    • Place of Presentation
      国立京都国際会館
    • Year and Date
      2011-09-24
    • Related Report
      2011 Final Research Report
  • [Presentation] 亜鉛Phos-tag SDS-PAGEによる細胞内キナーゼ/フォスファターゼの動態解析2011

    • Author(s)
      木下恵美子
    • Organizer
      第84回日本生化学会
    • Place of Presentation
      京都市
    • Year and Date
      2011-09-24
    • Related Report
      2011 Annual Research Report
  • [Presentation] 改良型Phos-tag Biotinを用いたペプチドマイクロアレイ解析による蛋白質リン酸化反応プロファイリング2011

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第9回日本プロテオーム学会大会
    • Place of Presentation
      朱鷺メッセ,新潟
    • Year and Date
      2011-07-29
    • Related Report
      2011 Final Research Report
  • [Presentation] hos-tag技術の開発とリン酸化プロテオミクスへの応用2011

    • Author(s)
      木木下恵美子
    • Organizer
      第9回日本プロテオーム学会大会
    • Place of Presentation
      朱鷺メッセ,新潟
    • Year and Date
      2011-07-29
    • Related Report
      2011 Final Research Report
  • [Presentation] 中性ゲルを用いた改良型Phos-tag SDS-PAGEによるタンパク質リン酸化フォームの詳細な分離解析法2011

    • Author(s)
      木下恵美子, 木下英司, 小池透
    • Organizer
      第9回日本プロテオーム学会
    • Place of Presentation
      新潟市
    • Year and Date
      2011-07-29
    • Related Report
      2011 Annual Research Report
  • [Presentation] 改良型Phos-tag Biotinを用いたペプチドマイクロアレイ解析による蛋白質リン酸化反応プロファイリング2011

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第9回日本プロテオーム学会
    • Place of Presentation
      新潟市
    • Year and Date
      2011-07-29
    • Related Report
      2011 Annual Research Report
  • [Presentation] Phos-tag技術の開発とリン酸化プロテオミクスへの応用2011

    • Author(s)
      木下恵美子
    • Organizer
      第9回日本プロテオーム学会
    • Place of Presentation
      新潟市(招待講演)
    • Year and Date
      2011-07-29
    • Related Report
      2011 Annual Research Report
  • [Presentation] 中性ゲルを用いた改良型Phos-tag SDS-PAGEによるタンパク質リン酸化フォームの詳細な分離解析法2011

    • Author(s)
      木下恵美子, 木下英司, 小池透
    • Organizer
      第9回日本プロテオーム学会大会
    • Place of Presentation
      朱鷺メッセ,新潟
    • Year and Date
      2011-07-28
    • Related Report
      2011 Final Research Report
  • [Presentation] Phos-tag-based fluorescence resonanceenergy transfer system for the analysis of phosphorylated molecules2010

    • Author(s)
      Masao Somura, Kei Takiyama, TomomiNagai, Eiji Kinoshita, EmikoKinoshita-Kikuta, and Tohru Koike
    • Organizer
      2010International Chemical Congress of PacificBasin Societies
    • Place of Presentation
      Kamehameha Halls, Honolulu, Hawaii, USA.
    • Year and Date
      2010-12-16
    • Related Report
      2011 Final Research Report
  • [Presentation] Quantitative analysis of the phosphorylation states in a bacterialtwo-component system using Phos-tag SDS-PAGE2010

    • Author(s)
      Shiho Yanagihara, Emiko Kinoshita-Kikuta, Eiji Kinoshita, and Tohru Koike
    • Organizer
      2010International ChemicalCongress of Pacific Basin Societies
    • Place of Presentation
      Kamehameha Halls, Honolulu, Hawaii
    • Year and Date
      2010-12-16
    • Related Report
      2011 Final Research Report
  • [Presentation] Phos-tagSDS-PAGE for protein phosphorylationprofiling2010

    • Author(s)
      Eiji Kinoshita, Emiko Kinoshita-Kikuta, Saori Okatani, and Tohru Koike
    • Organizer
      2010International ChemicalCongress of Pacific Basin Societies
    • Place of Presentation
      Kamehameha Halls, Honolulu, Hawaii, USA
    • Year and Date
      2010-12-16
    • Related Report
      2011 Final Research Report
  • [Presentation] Phos-tag SDS-PAGE for protein phosphorylation profiling2010

    • Author(s)
      Kinoshita, E.
    • Organizer
      2010 International Chemical Congress of Pacific Basin Societies
    • Place of Presentation
      Convention Center, Honolulu, Hawaii, USA
    • Year and Date
      2010-12-16
    • Related Report
      2010 Annual Research Report
  • [Presentation] Quantitative analysis of the phosphorylation states in a bacterial two-component system using Phos-tag SDS-PAGE2010

    • Author(s)
      Yanagihara, S.
    • Organizer
      2010 International Chemical Congress of Pacific Basin Societies
    • Place of Presentation
      Convention Center, Honolulu, Hawaii, USA
    • Year and Date
      2010-12-16
    • Related Report
      2010 Annual Research Report
  • [Presentation] A Phos-tag-basedfluorescence resonance energy transfersystem for the analysis of the kinasereaction of a substrate peptide2010

    • Author(s)
      Masao Somura, Kei Takiyama, EmikoKinoshita-Kikuta, Eiji Kinoshita, andTohru Koike
    • Organizer
      第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
    • Place of Presentation
      神戸国際展示場
    • Year and Date
      2010-12-09
    • Related Report
      2011 Final Research Report
  • [Presentation] Improved Phos-tag Biotin for advanced profiling of proteinphosphorylation2010

    • Author(s)
      Eiji Kinoshita, Emiko Kinoshita-Kikuta, and Tohru Koike
    • Organizer
      第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
    • Place of Presentation
      神戸国際展示場
    • Year and Date
      2010-12-09
    • Related Report
      2011 Final Research Report
  • [Presentation] Improved Phos-tag SDS-PAGE under neutral pH conditionsfor advanced protein phosphorylationprofiling2010

    • Author(s)
      Emiko Kinoshita-Kikuta, Eiji Kinoshita, and Tohru Koike
    • Organizer
      第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
    • Place of Presentation
      神戸国際展示場
    • Year and Date
      2010-12-09
    • Related Report
      2011 Final Research Report
  • [Presentation] Improved Phos-tag SDS-PAGE under neutral pH conditions for advanced protein phosphorylation profiling2010

    • Author(s)
      木下恵美子
    • Organizer
      第33回日本分子生物学会・第83回日本生化学学会年会
    • Place of Presentation
      神戸国際展示場
    • Year and Date
      2010-12-09
    • Related Report
      2010 Annual Research Report
  • [Presentation] 高分子量リン酸化蛋白質解析のための改良型Phos-tag電気泳動法2010

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第61回日本電気泳動学会総会
    • Place of Presentation
      北海道大学
    • Year and Date
      2010-09-19
    • Related Report
      2011 Final Research Report
  • [Presentation] 蛋白質リン酸化反応を高感度検出するための改良型Phos-tag Biotin2010

    • Author(s)
      木下英司, 木下恵美子, 小池透
    • Organizer
      第8回日本プロテオーム学会大会
    • Place of Presentation
      東京ベイホテル東急
    • Year and Date
      2010-07-27
    • Related Report
      2011 Final Research Report
  • [Presentation] Improved Phos-tag Biotin for highly sensitive detection of protein phosphorylation2010

    • Author(s)
      木下英司
    • Organizer
      日本プロテオーム学会第8回大会
    • Place of Presentation
      東京ベイホテル東急
    • Year and Date
      2010-07-27
    • Related Report
      2010 Annual Research Report
  • [Presentation] 新規リン酸親和性Phos-tagビーズを用いたヌクレオチド類の分離精製法2010

    • Author(s)
      宗村雅男, 木下英司, 木下恵美子, 山田篤志, 小池透
    • Organizer
      第51回日本生化学会中国・四国支部例会
    • Place of Presentation
      山口大学
    • Year and Date
      2010-05-14
    • Related Report
      2011 Final Research Report
  • [Presentation] 新規リン酸親和性Phos-tagビーズを用いたリン酸化タンパク質の分離濃縮法2010

    • Author(s)
      木下英司, 木下恵美子, 山田篤志, 小池透
    • Organizer
      第51回日本生化学会中国・四国支部例会
    • Place of Presentation
      山口大学
    • Year and Date
      2010-05-14
    • Related Report
      2011 Final Research Report
  • [Remarks]

    • URL

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

    • Related Report
      2011 Final Research Report
  • [Remarks]

    • URL

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

    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

      http://www.phos-tag.com/

    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

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

    • Related Report
      2010 Annual Research Report

URL: 

Published: 2010-08-23   Modified: 2016-04-21  

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