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Identification of endogenous kinase substrates for analyzing intracellular signaling pathway

Research Project

Project/Area Number 17H03605
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Genome biology
Research InstitutionKyoto University

Principal Investigator

Ishihama Yasushi  京都大学, 薬学研究科, 教授 (30439244)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2019: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2017: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Keywordsキナーゼ / キナーゼ基質 / リン酸化プロテオーム / 細胞内シグナル伝達 / リン酸化シグナル / プロテオーム
Outline of Final Research Achievements

The goal of this study is to establish a method to identify the endogenous kinase substrates on a large scale, and to apply to intracellular signal pathway analysis. To accomplish this goal, we have developed three methods as follows: (1) a method to measure changes in phosphorylation upon kinase perturbation, (2) a method to calculate the motif score based on the extracted phosphorylation motif of each kinase from in vitro kinase assay, and (3) a method to analyze the kinase-substrate transient interactions. These three methods were combined and applied to identifying endogenous kinase substrates successfully. In addition, we developed a method for estimating the activated kinase from the phosphoproteome information based on kinase-substrate relationships.

Academic Significance and Societal Importance of the Research Achievements

国内外のゲノムワイド関連解析により、様々な疾病原因は個々の遺伝子よりもシグナルパスウェイ毎の異常に集積されており、特に癌は十数種のパスウェイに原因遺伝子が分布することがわかってきた。これらのパスウェイにはすべてリン酸化シグナルが関わっており、その異常はガンそのものの発症因子ともなり、その進行、薬剤感受性および治療予後とも深く関わる。本研究の成果により、様々なパスウェイ毎のリン酸化動態を全体として理解することが可能となり、がんをはじめとする疾病の分子基盤の解明につながるものと期待される。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (57 results)

All 2020 2019 2018 2017 Other

All Int'l Joint Research (2 results) Journal Article (6 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 6 results,  Open Access: 3 results) Presentation (49 results) (of which Int'l Joint Research: 22 results,  Invited: 15 results)

  • [Int'l Joint Research] University of California, San Francisco(米国)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] University of Manitoba(カナダ)

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Selected Reaction Monitoring of Kinase Activity-Targeted Phosphopeptides2019

    • Author(s)
      Chisato TAKAHASHI, Tatsuya YAZAKI, Naoyuki SUGIYAMA, Yasushi ISHIHAMA
    • Journal Title

      CHROMATOGRAPHY

      Volume: 40 Issue: 2 Pages: 39-47

    • DOI

      10.15583/jpchrom.2019.005

    • NAID

      130007676149

    • ISSN
      1342-8284, 1348-3315
    • Year and Date
      2019-06-20
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Large-scale Discovery of Substrates of the Human Kinome2019

    • Author(s)
      Sugiyama Naoyuki、Imamura Haruna、Ishihama Yasushi
    • Journal Title

      Scientific Reports

      Volume: 9 Issue: 1 Pages: 10503-10503

    • DOI

      10.1038/s41598-019-46385-4

    • NAID

      120006726156

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Mitochondrial lipoylation integrates age-associated decline in brown fat thermogenesis2019

    • Author(s)
      Tajima Kazuki、Ikeda Kenji、Chang Hsin-Yi、Chang Chih-Hsiang、Yoneshiro Takeshi、Oguri Yasuo、Jun Heejin、Wu Jun、Ishihama Yasushi、Kajimura Shingo
    • Journal Title

      Nature Metabolism

      Volume: 1 Issue: 9 Pages: 886-898

    • DOI

      10.1038/s42255-019-0106-z

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Comparative proteomics of Helicobacter pylori strains reveals geographical features rather than genomic variations2019

    • Author(s)
      Sugiyama Naoyuki、Miyake Satomi、Lin Miao‐Hsia、Wakabayashi Masaki、Marusawa Hiroyuki、Nishiumi Shin、Yoshida Masaru、Ishihama Yasushi
    • Journal Title

      Genes to Cells

      Volume: 24 Issue: 2 Pages: 139-150

    • DOI

      10.1111/gtc.12662

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Retention Order Reversal of Phosphorylated and Unphosphorylated Peptides in Reversed-Phase LC/MS2018

    • Author(s)
      OGATA Kosuke、KROKHIN Oleg V.、ISHIHAMA Yasushi
    • Journal Title

      Analytical Sciences

      Volume: 34 Issue: 9 Pages: 1037-1041

    • DOI

      10.2116/analsci.18SCP11

    • NAID

      130007474696

    • ISSN
      0910-6340, 1348-2246
    • Year and Date
      2018-09-10
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Absolute Phosphorylation Stoichiometry Analysis by Motif-Targeting Quantitative Mass Spectrometry2017

    • Author(s)
      Tsai Chia-Feng、Ku Wei-Chi、Chen Yu-Ju、Ishihama Yasushi
    • Journal Title

      Methods in Molecular Biology

      Volume: 1636 Pages: 313-325

    • DOI

      10.1007/978-1-4939-7154-1_20

    • ISBN
      9781493971527, 9781493971541
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] Challenges to illuminate human kinome2020

    • Author(s)
      Yasushi Ishihama
    • Organizer
      AOMSC2020
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Comprehensive monitoring of intracellular tyrosine kinase activities2020

    • Author(s)
      Kazutaka Yachigo, Dai Sakamoto, Tatsuya Yazaki, Chisato Takahashi, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      4th Symposium on Kyoto Biomolecular Mass Spectrometry Society
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Kinome-wide Libraries of Exogenous Substrates for Profiling Intracellular Kinome Activities2020

    • Author(s)
      Junna Nakazono, Dai Sakamoto, Naoyuki Sugiyama and Yasushi Ishihama
    • Organizer
      4th Symposium on Kyoto Biomolecular Mass Spectrometry Society
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 配位子交換クロマトグラフィーによるリン酸化チロシン選択的ペプチド濃縮法の開発2020

    • Author(s)
      中田花菜、南部早紀、杉山直幸、石濱泰
    • Organizer
      日本薬学会第140年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Toolbox for Phosphoproteomics in Cancer Signaling2019

    • Author(s)
      Yasushi Ishihama
    • Organizer
      Keystone Symposia_Proteomics and its Application to Translational and Precision Medicine
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Additional Separation Space by Trapped Ion Mobility Spectrometry Improves Accuracy of Isobaric Tag-based Quantitation2019

    • Author(s)
      Kosuke Ogata, Tomoya Niinae, Chia-Feng Tsai, Koshi Imami, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      Keystone Symposia_Proteomics and its Application to Translational and Precision Medicine
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 人工基質ペプチドを用いたキノーム活性プロファイリング2019

    • Author(s)
      杉山直幸、中園純菜、八児一隆、坂本大、Hsin-Yi Chang、石濱泰
    • Organizer
      第67回質量分析総合討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Proximity-dependent identification of in vivo substrates of putative kinases2019

    • Author(s)
      Tomoya Niinae, Koshi Imami, Chia-Feng Tsai, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      第67回アメリカ質量分析学会(ASMS2019)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Kinome Activity Analysis Based on Quantitative Phosphoproteomics2019

    • Author(s)
      Yuki Yoshikawa, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      日本プロテオーム学会2019年大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Development of novel method for identifying kinase endogenous substrates and application to mutant substrates analysis2019

    • Author(s)
      Tomoya Niinae, Koshi Imami, Chia-Feng Tsai, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      日本プロテオーム学会2019年大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] キナーゼ基質ペプチドライブラリを用いた細胞内チロシンキナーゼ活性計測2019

    • Author(s)
      八児一隆、坂本大、高橋知里、杉山直幸、石濱泰
    • Organizer
      BMAS2019 第32回バイオメディカル分析科学シンポジウム
    • Related Report
      2019 Annual Research Report
  • [Presentation] プロテオミクスLC/MSにおけるリン酸化ペプチドの保持挙動2019

    • Author(s)
      小形公亮、杉山直幸、KROKHIN Oleg V.、石濱泰
    • Organizer
      PPF2019 第17回次世代を担う若手のためのフィジカル・ファーマフォーラム
    • Related Report
      2019 Annual Research Report
  • [Presentation] Toolbox for human proteome, kinome and phosphatome analysis2019

    • Author(s)
      Yasushi Ishihama
    • Organizer
      Short course in Frontier Sciences and Technology: Data Analysis in Quantitative Proteomics
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] セリン/スレオニンキナーゼ特異的人工基質ペプチドを用いたキノーム活性計測2019

    • Author(s)
      中園 純菜・坂本 大・杉山 直幸・石濱 泰
    • Organizer
      日本分析化学会 第68年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Proximity-dependent approach for identifying putative endogenous substrates of protein kinases2019

    • Author(s)
      Tomoya Niinae・Koshi Imami・Chia-Feng Tsai・Naoyuki Sugiyama・Yasushi Ishihama
    • Organizer
      HUPO2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] In vitro Profiling of Ser/Thr/Tyr Selectivity of Human Protein Kinome2019

    • Author(s)
      Naoyuki Sugiyama, Kazutaka Yachigo, Masato Tsuyuguchi, Takayoshi Kinoshita, Yasushi Ishihama
    • Organizer
      HUPO2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Monitoring intracellular kinome activities using kinase specific substrate peptides2019

    • Author(s)
      Junna Nakazono, Dai Sakamoto, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      HPLC2019 Kyoto
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Targeted phosphoproteomics for profiling tyrosine kinase activities2019

    • Author(s)
      Kazutaka Yachigo, Tatsuya Yazaki, Chisato Takahashi, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      HPLC2019 Kyoto
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] キナーゼ特異的人工基質ライブラリを用いた細胞内キノーム活性プロファイリング2019

    • Author(s)
      中園 純菜, 坂本 大, 杉山 直幸, 石濱 泰
    • Organizer
      第30回クロマトグラフィー科学会議 (SCS30)
    • Related Report
      2019 Annual Research Report
  • [Presentation] ターゲットプロテオミクスによるチロシンキナーゼ活性計測2019

    • Author(s)
      八児一隆、矢崎達也、高橋知里、杉山直幸、石濱泰
    • Organizer
      第30回クロマトグラフィー科学会議 (SCS30)
    • Related Report
      2019 Annual Research Report
  • [Presentation] Evaluation of kinase substrate relationships2019

    • Author(s)
      Tomoya Niinae, Koshi Imami, Chia-Feng Tsai, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      3rd KBMSSシンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] キナーゼ収斂リン酸化プロテオミクスを用いた分子標的薬プロファイリング2019

    • Author(s)
      吉川侑樹・杉山直幸・石濱泰
    • Organizer
      日本薬学会第139年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] キナーゼ基質大規模同定に基づくキナーゼの基質配列指向性評価2018

    • Author(s)
      中園純菜・ 坂本大・ 張心儀・ 杉山直幸・ 石濱泰
    • Organizer
      Mass Spectrometry and Proteomics 2018 (MSP2018)(日本質量分析学会・日本プロテオーム学会 2018年合同大会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] チロシンキナーゼ特異的人工基質ペプチドを用いたin vitroキノーム活性計測2018

    • Author(s)
      八児一隆・ 高橋知里・ 坂本大・ 張心儀・ 杉山直幸・ 石濱泰
    • Organizer
      Mass Spectrometry and Proteomics 2018 (MSP2018)(日本質量分析学会・日本プロテオーム学会 2018年合同大会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] キノームプロファイリングによるリン酸化ネットワーク解析2018

    • Author(s)
      杉山直幸・ 今村春菜・ 坂本大・ 中園純菜・ 八児一隆・ 高橋知里・ Hsin-yi, Chang・ 石濱泰
    • Organizer
      Mass Spectrometry and Proteomics 2018 (MSP2018)(日本質量分析学会・日本プロテオーム学会 2018年合同大会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] H. pylori proteomics2018

    • Author(s)
      Naoyuki Sugiyama
    • Organizer
      International Proteogenome Workshop in Kyoto
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 固定化金属イオンアフィニティークロマトグラフィーを用いたチロシンリン酸化ペプチド濃縮法の開発2018

    • Author(s)
      南部早紀、小笠原実穂、張心儀、杉山直幸、石濱泰
    • Organizer
      BMAS2018
    • Related Report
      2018 Annual Research Report
  • [Presentation] Direct Profiling of Kinome Activities Using Kinase-Specific Substrate Peptides2018

    • Author(s)
      "Naoyuki Sugiyama, Dai Sakamoto, Junna Nakazono, Kazutaka Yachigo, Hsin-Yi Chang, Yasushi Ishihama"
    • Organizer
      25th International Symposium on Electro- and Liquid Phase-Separation Techniques(ITP2018)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Phosphoproteome and Kinome Profiling Using NanoLC-MS/MS2018

    • Author(s)
      Naoyuki Sugiyama
    • Organizer
      2nd International BMS Symposium 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 固定化金属イオンに対する相互作用に基づいたチロシンリン酸化ペプチド濃縮法の開発2018

    • Author(s)
      南部早紀、小笠原実穂、張心儀、杉山直幸、石濱泰
    • Organizer
      第29回クロマトグラフィー科学会議
    • Related Report
      2018 Annual Research Report
  • [Presentation] Human Kinome Profiling by Quantitative Phosphoproteomics2018

    • Author(s)
      Yasushi Ishihama
    • Organizer
      2018 Annual meeting of Taiwan Proteomics Society
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Human Kinome and Phosphatome Profiling2018

    • Author(s)
      Yasushi Ishihama
    • Organizer
      The 9th International Forum on Chemistry of Functional Organic Chemicals (IFOC-9)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] プロテオミクスによる細胞内キナーゼ基質大規模同定法の開発2018

    • Author(s)
      新苗 智也, 今見 考志, Tsai Chia-Feng, 杉山 直幸, 石濱 泰
    • Organizer
      第41回日本分子生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] キノーム活性測定に向けたin vitroキナーゼ基質の大規模同定2018

    • Author(s)
      中園純菜・ 坂本大・ 張心儀・ 杉山直幸・ 石濱泰
    • Organizer
      第41回日本分子生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] セリウム錯体を用いた脱リン酸化法のin vitroキナーゼ試験への適用2018

    • Author(s)
      八児 一隆・ 張 心儀・ 杉山 直幸・ 石濱 泰
    • Organizer
      第41回日本分子生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] キナーゼ収斂型リン酸化プロテオミクス2018

    • Author(s)
      石濱 泰
    • Organizer
      次世代脳プロジェクト冬のシンポジウム若手ワークショップ「トランスオミクスによる精神疾患の分子基盤解明に向けて」
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] Tyrosine phosphoproteomics based on immobilized metal ion affinity chromatography2018

    • Author(s)
      " Saki Nambu, Miho Ogasawara, Hsin-Yi Chang, Naoyuki Sugiyama, Yasushi Ishihama "
    • Organizer
      3rd KBMSSシンポジウム
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Human Kinome Profiling using Phosphoproteomic Approaches2018

    • Author(s)
      Naoyuki Sugiyama , Yasushi Ishihama
    • Organizer
      KBMSS2018
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] ショットガンプロテオミクスにおける計測と情報の新展開2018

    • Author(s)
      石濱 泰
    • Organizer
      第15回北里疾患プロテオーム研究会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 細胞抽出タンパク質プールを用いたキナーゼの基質モチーフ解析2018

    • Author(s)
      新苗智也・今見考志・Hsin-Yi Chang・杉山直幸・石濱泰
    • Organizer
      日本薬学会第138年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] キナーゼ阻害薬と定量的リン酸化プロテオミクスを用いたPKA基質探索2017

    • Author(s)
      新苗智也・小形公亮・今村春菜・若林真樹・杉山直幸・石濱泰
    • Organizer
      第65回質量分析総合討論会
    • Related Report
      2017 Annual Research Report
  • [Presentation] リン酸化プロテオーム解析技術を用いたキノームプロファイリング2017

    • Author(s)
      杉山直幸、石濱 泰
    • Organizer
      第77回分析化学討論会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Kinase activity profiling by motif-targeting phosphoproteome analysis2017

    • Author(s)
      Chia-Feng Tsai, Masaki Wakabayashi, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      65th ASMS
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] プロテオミクスを用いたキナーゼ収斂型シグナルネットワーク解析2017

    • Author(s)
      杉山直幸、石濱 泰
    • Organizer
      質量分析フォーラム2017
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Kinase activity profiling by motif-targeting phosphoproteome analysis2017

    • Author(s)
      Chia-Feng Tsai, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      日本プロテオーム学会2017年大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 異なる細胞株由来タンパク質に対するキナーゼのリン酸化モチーフ解析2017

    • Author(s)
      新苗智也、張心儀、杉山直幸、石濱泰
    • Organizer
      第28回クロマトグラフィー科学会議
    • Related Report
      2017 Annual Research Report
  • [Presentation] プロテオミクスを用いて暗黒キノーム世界を照らす2017

    • Author(s)
      石濱 泰
    • Organizer
      第37回キャピラリー電気泳動シンポジウム
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] プロテオミクスを用いたキノーム活性プロファイリング2017

    • Author(s)
      石濱 泰
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Kinome activity profiling based on large-scale kinase-substrate relationships2017

    • Author(s)
      Yasushi Ishihama
    • Organizer
      AOMSC 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited

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Published: 2017-04-28   Modified: 2021-02-19  

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