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Development of Novel Fluorogenic Probes for Labeling Intracellular Proteins

Research Project

Project/Area Number 22685016
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Chemistry related to living body
Research InstitutionOsaka University

Principal Investigator

HORI Yuichiro  大阪大学, 工学(系)研究科(研究院), 助教 (00444563)

Project Period (FY) 2010-04-01 – 2013-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥24,570,000 (Direct Cost: ¥18,900,000、Indirect Cost: ¥5,670,000)
Fiscal Year 2012: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2011: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
Fiscal Year 2010: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
Keywords蛋白質標識 / 生細胞蛍光イメージング / 発蛍光プローブ / PYPタグ / タグ蛋白質 / 無洗浄イメージング / 蛋白質標識法 / 蛍光プローブ / PYP / 蛍光強度増大 / 桂皮酸誘導体 / クマリン誘導体
Research Abstract

Fluorescence protein labeling by a specific pair of a protein tag and synthetic probe is a powerful chemistry-based approach to investigate protein function and localization inside living cells. In this research, we developed a protein labeling system based on fluorogenic probes and photoactive yellow protein (PYP) tag. By utilizing this technique, we were succeeded in specific imaging of proteins inside living cells with high S/N ratio and excellent labeling kinetics. Furthermore, the labeling technique was applied to the imaging of DNA methylation.

Report

(4 results)
  • 2013 Final Research Report ( PDF )
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (31 results)

All 2014 2013 2012 2011 2010 Other

All Journal Article (12 results) (of which Peer Reviewed: 12 results,  Open Access: 1 results) Presentation (17 results) (of which Invited: 2 results) Remarks (1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Development of Fluorogenic Probes for Quick No-Wash Live-Cell Imaging of Intracellular Proteins2013

    • Author(s)
      Y. Hori, T. Norinobu, M. Sato, K. Arita, S. Shirakawa & K. Kikuchi
    • Journal Title

      J. Am. Chem. Soc

      Volume: 135 Issue: 33 Pages: 12360-12365

    • DOI

      10.1021/ja405745v

    • Related Report
      2013 Final Research Report 2012 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Protein Labeling with Fluorogenic Probes for No-Wash Live-Cell Imaging of Proteins2013

    • Author(s)
      Hori, Y., Kikuchi, K.
    • Journal Title

      Curr. Opin. Chem. Biol.

      Volume: 17 Issue: 4 Pages: 644-650

    • DOI

      10.1016/j.cbpa.2013.05.015

    • Related Report
      2013 Final Research Report 2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 発蛍光型スイッチ機能を有した蛋白質ラベル化技術の開発による細胞内標的分子イメージング2013

    • Author(s)
      堀 雄一郎、菊地 和也
    • Journal Title

      CSJ Current Review

      Volume: 12 Pages: 162-168

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] 発蛍光型スイッチ機能を有した蛋白質ラベル化技術の開発による細胞内標的分子イメージング2013

    • Author(s)
      堀 雄一郎、菊地 和也
    • Journal Title

      化学と生物

      Volume: 51 Pages: 189-192

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Development of Protein-Labeling Probes with Redesigned Fluorogenic Switch Based on Intramolecular Association for No-wash Live-cell Imaging2012

    • Author(s)
      Hori, Y., Nakaki, K., Sato, M., Mizukami, S., Kikuchi, K.
    • Journal Title

      Angew. Chem. Int. Ed

      Volume: (In press) Issue: 23 Pages: 5611-5614

    • DOI

      10.1002/anie.201200867

    • Related Report
      2013 Final Research Report 2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Development of Fluorogenic Probe with Transesterification Switch for Detection of Histone Deacetylase Activity2012

    • Author(s)
      R. Baba, Y. Hori, S. Mizukami & K. Kikuchi
    • Journal Title

      J. Am. Chem. Soc

      Volume: 134 Issue: 35 Pages: 14310-14313

    • DOI

      10.1021/ja306045j

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Fluorescent Probe for Detection of His tone Deacetylase Activity Based on Aggregation-Induced Emission2012

    • Author(s)
      Dhara, K., Hori, Y., Baba, R, Kikuchi, K.
    • Journal Title

      Chem. Commun.

      Volume: 48 Issue: 94 Pages: 11534-11536

    • DOI

      10.1039/c2cc36591j

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] 発蛍光プローブによる次世代型タンパク質標識技術-化学アプローチに基づくタンパク質蛍光イメージングの新展開2012

    • Author(s)
      堀 雄一郎、菊地 和也
    • Journal Title

      化学

      Volume: 67 Pages: 68-69

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] 合成蛍光プローブと発現タンパク質を用いた新規タンパク質ラベル化システム2011

    • Author(s)
      堀 雄一郎、菊地 和也
    • Journal Title

      生化学

      Volume: 83 Pages: 135-139

    • NAID

      10029067602

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] 合成蛍光プローブによるタンパク質ラベル化法の開発2011

    • Author(s)
      堀雄一郎
    • Journal Title

      生化学

      Volume: 83 Pages: 135-139

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Multicolor Protein Labeling in Living Cells Using Mutant β-Lactamase-Tag Technology2010

    • Author(s)
      Shuji Watanabe
    • Journal Title

      Bioconjugate Chemistry

      Volume: 21 Pages: 2320-2326

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Photocontrolled Compound Release System Using Caged Antimicrobial Peptide2010

    • Author(s)
      Shin Mizukami
    • Journal Title

      Journal of the American Chemical Society

      Volume: 132 Pages: 9524-9525

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] Development of Small Molecules with Fluorogenic Switches for Visualizing Protein Function and Localization2014

    • Author(s)
      Hori, Y.
    • Organizer
      iCeMS-RIKEN Joint Symposium on Mesoscopic Chemical Biology Integrated Chemical-Physical Systems Towards Cell Control
    • Place of Presentation
      Kyoto, Japan
    • Year and Date
      2014-02-07
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] Development of PYP-tag mutants and fluorogenic probes for rapid imaging of intracellular proteins2013

    • Author(s)
      Hori, Y., Sato, M., Kikuchi, K.
    • Organizer
      Seeing is Believing, EMBO/EMBL Symposium
    • Place of Presentation
      Heidelberg, Germany
    • Related Report
      2013 Final Research Report
  • [Presentation] 環境応答性発蛍光プローブとPYP タグを利用した細胞内蛋白質高速イメージング技術の開発と生物応用2013

    • Author(s)
      堀雄一郎、佐藤基、菊地和也
    • Organizer
      第7回バイオ関連化学シンポジウム
    • Place of Presentation
      名古屋
    • Related Report
      2013 Final Research Report
  • [Presentation] 環境応答性発蛍光プローブとPYPタグを利用した細胞内蛋白質高速イメージング技術の開発と生物応用2013

    • Author(s)
      堀雄一郎, 佐藤基, 菊地和也
    • Organizer
      第7回バイオ関連化学シンポジウム
    • Place of Presentation
      名古屋
    • Related Report
      2012 Annual Research Report
  • [Presentation] Development of PYP-tag mutants and fluorogenic probes for rapid imaging of intracellular proteins2013

    • Author(s)
      Yuichiro Hori, Motoki Sato, Kazuya Kikuchi
    • Organizer
      EMBO/EMBL symposium, Seeing is Believing
    • Place of Presentation
      Heidelberg (Germany)
    • Related Report
      2012 Annual Research Report
  • [Presentation] Development of Proteinlabeling Probes with Environmentsensitive Fluorogenic Switch for No-wash Labeling of Intracellular Proteins Tagged with Photoactive Yellow Protein (PYP) Mutant2012

    • Author(s)
      Hori, Y., Sato, M., Kamikawa, Y., Kikuchi, K.
    • Organizer
      EMBL Conference Series, Chemical Biology 2012
    • Place of Presentation
      Heidelberg, Germany
    • Year and Date
      2012-09-27
    • Related Report
      2013 Final Research Report
  • [Presentation] Development of Fluorogenic Probes with Chemical Triggers for Imaging Protein Function and Localization2012

    • Author(s)
      Hori, Y.
    • Organizer
      The 5^<th> "MEXT Project of Integrated Research on Chemical Synthesis" Forum, Unique peptide assemblies with biological functions
    • Place of Presentation
      Uji, Japan
    • Year and Date
      2012-09-15
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] PYPタグと発蛍光プローブを利用した生細胞蛋白質イメージング2012

    • Author(s)
      堀 雄一郎、佐藤 基、菊地 和也
    • Organizer
      第3回Vivid Workshop
    • Place of Presentation
      加賀
    • Year and Date
      2012-02-21
    • Related Report
      2013 Final Research Report
  • [Presentation] Design of Fluorogenic Probes for Labeling Photoactive Yellow Protein (PYP) Tag and No-wash Live-cell Imaging2012

    • Author(s)
      Yuichiro Hori, Kyohei Nakaki, Tomoya Norinobu, Motoki Sato, Kazuya Kikuchi
    • Organizer
      Gordon Research Conference
    • Place of Presentation
      Andover (USA)
    • Related Report
      2012 Annual Research Report
  • [Presentation] Development of Photoactive Yellow Protein-based Protein Labeling System with Designed Fluorescence Probes2011

    • Author(s)
      Hori, Y., Nakaki, K., Norinobu, T., Kikuchi, K.
    • Organizer
      Seeing is Believing, EMBO/EMBL Symposium
    • Year and Date
      2011-03-18
    • Related Report
      2013 Final Research Report
  • [Presentation] Design of Fluorogenic Probes for Protein Labeling Method Based on Photoactive yellow Protein (PYP) Tag and Live Cell Imaging2011

    • Author(s)
      Hori, Y., Kikuchi, K.
    • Organizer
      Nature Chemical Biology Symposium
    • Place of Presentation
      Boston, USA
    • Related Report
      2013 Final Research Report
  • [Presentation] hotoactive Yellow Protein(PYP)タグと蛍光強度増大型プローブを利用した蛋白質標識法の開発と生細胞イメージング2011

    • Author(s)
      堀雄一郎、中木恭兵、則信智哉、菊地和也
    • Organizer
      バイオ関連化学シンポジウム
    • Place of Presentation
      筑波
    • Related Report
      2013 Final Research Report
  • [Presentation] Photoactive Yellow Protein(PYP)タグと蛍光強度増大型プローブを利用した蛋白質標識法の開発と生細胞イメージング2011

    • Author(s)
      堀雄一郎、中木恭兵、則信智也、菊地和也
    • Organizer
      5回バイオ関連化学シンポジウム
    • Place of Presentation
      つくば(茨城)
    • Related Report
      2011 Annual Research Report
  • [Presentation] Design of fluorogenic probes for protein labeling method based on photoactive yellow protein (PYP) tag and live cell imaging2011

    • Author(s)
      Hori, Y., Nakaki, K., Norinobu, T., Kikuchi, K.
    • Organizer
      Nature Chemical Biology Symposium 2011: Cancer Chemical Biology
    • Place of Presentation
      Boston (USA)
    • Related Report
      2011 Annual Research Report
  • [Presentation] Development of a Protein Labeling System Based on Photoactive Yellow Protein and a Designed Fluorogenic Probe2010

    • Author(s)
      Hori, Y., Nakaki, K., Kikuchi, K.
    • Organizer
      Pacifichem 2010
    • Place of Presentation
      Honolulu, USA
    • Year and Date
      2010-12-19
    • Related Report
      2013 Final Research Report
  • [Presentation] Design of Fluorogenic Probe for Novel Labeling System Based on Protein Tag, Photoactive Yellow Protein2010

    • Author(s)
      堀 雄一郎、則信 智也、中木 恭兵、菊地 和也
    • Organizer
      BMB2010
    • Place of Presentation
      神戸
    • Year and Date
      2010-12-09
    • Related Report
      2013 Final Research Report
  • [Presentation] PYPタグと新規桂皮酸型プローブを利用した蛋白質ラベル化法の開発と無洗浄生細胞イメージング

    • Author(s)
      堀雄一郎、中木恭兵、佐藤基、菊地和也
    • Organizer
      日本化学会第92春季年会
    • Place of Presentation
      横浜(神奈川)
    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

      http://www-molpro.mls.eng.osaka-u.ac.jp

    • Related Report
      2013 Final Research Report
  • [Patent(Industrial Property Rights)] タンパク質を蛍光標識する方法2013

    • Inventor(s)
      菊地 和也,堀 雄一郎,上野 秀樹
    • Industrial Property Rights Holder
      国立大学法人大阪大学
    • Industrial Property Rights Type
      特許
    • Acquisition Date
      2013-06-28
    • Related Report
      2013 Final Research Report

URL: 

Published: 2010-08-23   Modified: 2019-07-29  

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