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Development of a system for profiling the pathophysiology of diseases from the phosphorylated states of proteins

Research Project

Project/Area Number 18K06596
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
Research InstitutionHiroshima University

Principal Investigator

Kinoshita Emiko  広島大学, 医系科学研究科(薬), 助教 (40379912)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
KeywordsPhos-tag / キナーゼ / リン酸化 / シグナル伝達 / phos-tag / フォスタグ / リン酸化タンパク質 / 疾患関連タンパク質
Outline of Final Research Achievements

In this study, we analyzed the relationship between disease-related kinase mutations and phosphorylation status using Phos-tag electrophoresis.Point mutations in MEK1 kinase are associated with the malignancy of certain sporadic cancer. Moreover, point mutations associated with anticancer drug resistance have also been reported. In this study, Phos-tag analysis was performed on the intracellular phosphorylation status of 15 types of MEK1 point mutations.As a result, it was shown that Phos-tag analysis is effective for the estimation of correlation between cancer and phosphorylation status, prediction of changes in kinase activity ,and prediction of drug efficacy.
In addition, we have set up a site to publish Phos-tag electrophoresis analysis data of proteins accumulated in previous studies (http://phostag.hiroshima-u.ac.jp).

Academic Significance and Societal Importance of the Research Achievements

ガンに関わるキナーゼの点変異が細胞内でのリン酸化状態にどのような変化をもたらすかをPhos-tag電気泳動方をもちいて調べた。この電気泳動パターンは,ガンの悪性度とリン酸化状態の相関,キナーゼ活性の変化や薬剤の有効性の予測などに役立つデータである。これまでにも,このようなPhos-tag電気泳動パターンを多く蓄積してきたので,本研究期間中にそれらのデータを公開するためのWebサイトを開設し,研究者に閲覧していただけるように整備している。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (23 results)

All 2021 2020 2019 2018 Other

All Journal Article (9 results) (of which Peer Reviewed: 9 results,  Open Access: 9 results) Presentation (12 results) Book (1 results) Remarks (1 results)

  • [Journal Article] An immuno-dot blot assay for screening histidine kinase inhibitors2020

    • Author(s)
      Kinoshita-Kikuta Emiko、Maruta Shino、Eguchi Yoko、Igarashi Masayuki、Okajima Toshihide、Utsumi Ryutaro、Kinoshita Eiji、Koike Tohru
    • Journal Title

      Analytical Biochemistry

      Volume: 600 Pages: 113765-113765

    • DOI

      10.1016/j.ab.2020.113765

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A dot-blot-staining method for detecting phosphoproteins with a Phos-tag Aqua fluorescent dye2020

    • Author(s)
      Kinoshita-Kikuta Emiko、Akayama Keisuke、Kinoshita Eiji、Koike Tohru
    • Journal Title

      Journal of Electrophoresis

      Volume: 64 Issue: 1 Pages: 7-11

    • DOI

      10.2198/jelectroph.64.7

    • NAID

      130007961315

    • ISSN
      1349-9394, 1349-9408
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Protein-N-myristoylation-dependent phosphorylation of serine 13 of tyrosine kinase Lyn by casein kinase 1γ at the Golgi during intracellular protein traffic2020

    • Author(s)
      Kinoshita-Kikuta Emiko、Utsumi Toshihiko、Miyazaki Aya、Tokumoto Chiharu、Doi Kyosuke、Harada Haruna、Kinoshita Eiji、Koike Tohru
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 16273-16273

    • DOI

      10.1038/s41598-020-73248-0

    • NAID

      120007169771

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Quantitative analysis of phosphoproteins in a bacterial two-component system using Phos-tag techniques2020

    • Author(s)
      Kinoshita-Kikuta Emiko、Kinoshita Eiji、Koike Tohru
    • Journal Title

      Electrophoresis Letters

      Volume: 64 Issue: 1 Pages: 35-39

    • DOI

      10.2198/electroph.64.35

    • NAID

      130007871304

    • ISSN
      2189-2628, 2189-2636
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A strategy to identify protein-N-myristoylation-dependent phosphorylation reactions of cellular proteins by using Phos-tag SDS-PAGE.2019

    • Author(s)
      Emiko Kinoshita-Kikuta, Ayane Tanikawa, Takuro Hosokawa, Aya Kiwado, Koko Moriya, Eiji Kinoshita, Tohru Koike, Toshihiko Utsumi
    • Journal Title

      PLOS ONE

      Volume: 14 Issue: 11 Pages: e0225510-e0225510

    • DOI

      10.1371/journal.pone.0225510

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Quantitative monitoring of His and Asp phosphorylation in a bacterial signaling system by using Phos-tag Magenta/Cyan fluorescent dyes2019

    • Author(s)
      Emiko Kinoshita-Kikuta, Hiroshi Kusamoto, Syogo Ono, Keisuke Akayama, Yoko Eguchi, Masayuki Igarashi, Toshihide Okajima, Ryutaro Utsumi, Eiji Kinoshita, Tohru Koike
    • Journal Title

      Electrophoresis

      Volume: 40 Issue: 22 Pages: 3005-3013

    • DOI

      10.1002/elps.201900261

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Gel-based analysis of protein phosphorylation status by rapid fluorometric staining using TAMRA-labeled Phos-tag2019

    • Author(s)
      Hiroshi Kusamoto, Emiko Kinoshita-Kikuta, Tomoyo Nishimura, Tomomi Nagai, Eiji Kinoshita, and Tohru Koike,
    • Journal Title

      Journal of Electrophoresis

      Volume: 63 Issue: 1 Pages: 25-32

    • DOI

      10.2198/jelectroph.63.25

    • NAID

      130007634033

    • ISSN
      1349-9394, 1349-9408
    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Increase in constitutively active MEK1 species by introduction of MEK1 mutations identified in cancers2019

    • Author(s)
      E. Kinoshita-Kikuta, E. Kinoshita, S. Ueda, Y. Ino, Y. Kimura, H. Hirano, and T. Koike
    • Journal Title

      BBA - Proteins and Proteomics

      Volume: 1867 Issue: 1 Pages: 62-70

    • DOI

      10.1016/j.bbapap.2018.05.004

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A method for profiling the phosphorylation state of tyrosine protein kinases.2018

    • Author(s)
      Yuuki Uezato, Isamu Kameshita, Keiko Morisawa, Shuji Sakamoto, Eiji Kinoshita, Emiko Kinoshita-Kikuta, Tohru Koike, Yasunori Sugiyama,
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

      Volume: 1867 Issue: 1 Pages: 71-75

    • DOI

      10.1016/j.bbapap.2018.05.003

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Phos-tag を用いたリン酸化タンパク質の解析2021

    • Author(s)
      木下恵美子
    • Organizer
      日本薬学会第141年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 細胞内のタンパク質のN-ミリストイル化に依存したリン酸化反応2020

    • Author(s)
      木下恵美子,木下英司,内海俊彦,小池透
    • Organizer
      第71回日本電気泳動学会総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 蛍光Phos-tag誘導体によるブロット膜上のリン酸化タンパク質の定量解析法の開発2020

    • Author(s)
      赤山圭佑,木下恵美子,木下英司,小池透
    • Organizer
      第93回日本生化学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Phos-tag 蛍光ゲル染色剤を用いた細菌の2成分伝達系のHis/Asp定量的リン酸化解析2020

    • Author(s)
      斧尚吾,木下恵美子,木下英司,小池透
    • Organizer
      第93回日本生化学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] チロシンキナーゼ Lyn は細胞膜への輸送の過程でゴルジ体においてcasein kinase 1γによってセリン13をリン酸化される2020

    • Author(s)
      木下恵美子,内海俊彦,木下英司、小池透
    • Organizer
      第93回日本生化学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Phos-tag を用いたリン酸化タンパク質の解析法2019

    • Author(s)
      木下恵美子
    • Organizer
      日本プロテオーム学会JPrOS2019,日本電気泳動学会JES2019合同年会
    • Related Report
      2019 Research-status Report
  • [Presentation] リン酸化タンパク質選択的な電気泳動ゲル蛍光染色法2019

    • Author(s)
      草本寛,木下恵美子,木下英司,小池透
    • Organizer
      日本プロテオーム学会JPrOS2019,日本電気泳動学会JES2019合同年会
    • Related Report
      2019 Research-status Report
  • [Presentation] 錯体医薬における亜鉛酵素阻害剤の親和性解析2019

    • Author(s)
      草本寛,木下恵美子,木下英司,小池透
    • Organizer
      医療薬学フォーラム2019/第27回クリニカルファーマシーシンポジウム
    • Related Report
      2019 Research-status Report
  • [Presentation] Phos-tag蛍光ゲル染色剤を用いた2成分系シグナリングの定量モニタリング2019

    • Author(s)
      木下英司,木下恵美子,江口陽子,五十嵐雅之,岡島俊英,内海龍太郎,小池 透
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Research-status Report
  • [Presentation] N-ミリストイル化依存的なタンパク質リン酸化反応のPhos-tag SDS-PAGEを用いた解析法の確立2018

    • Author(s)
      谷川 綾音、木下 恵美子、黄波戸 亜哉、守屋 康子、木下 英司、小池 透、内海 俊彦
    • Organizer
      第59回日本生化学会 中国四国支部例会
    • Related Report
      2018 Research-status Report
  • [Presentation] TAMRA-Phostagを用いたリン酸化タンパク質のゲル染色法2018

    • Author(s)
      草本寛,西村朋世,木下恵美子,木下英司,小池透
    • Organizer
      第69回日本電気泳動学会
    • Related Report
      2018 Research-status Report
  • [Presentation] TAMRA-Phostagを用いたリン酸化タンパク質のゲル染色法2018

    • Author(s)
      草本寛,西村朋世,木下恵美子,木下英司,小池透
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Research-status Report
  • [Book] Neuroproteomics: Neuromthods, 146 (ed. by Li, KW)2019

    • Author(s)
      Eiji Kinoshita, Kinoshita-Kikuta, E., Koike, T.
    • Total Pages
      218
    • Publisher
      Springer Nature
    • ISBN
      9781617791116
    • Related Report
      2019 Research-status Report
  • [Remarks] Phos-tag SDS-PAGE データ集

    • URL

      http://phostag.hiroshima-u.ac.jp

    • Related Report
      2020 Annual Research Report 2019 Research-status Report 2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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