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Shotgun proteomics beyond trypsin

Research Project

Project/Area Number 16K15107
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Physical pharmacy
Research InstitutionKyoto University

Principal Investigator

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

Co-Investigator(Renkei-kenkyūsha) SUGIYAMA Naoyuki  京都大学, 大学院薬学研究科, 准教授 (50545704)
WAKABAYASHI Masaki  京都大学, 大学院薬学研究科, 連携准教授 (70552024)
Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2016: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywordsショットガンプロテオミクス / トリプシン / トリプエヌ / タンパク質末端解析 / 分析化学 / プロテオミクス / 質量分析
Outline of Final Research Achievements

We investigated the usefulness of various proteases for shotgun proteomics, and found that TrypN, belonging to LysArginase family, has quite unique features. We optimized the digestion conditions as well as the separation conditions for tryp-N digested peptides. As a result, we found that tryp-N peptides can be separated based on the positive charged positions on peptides by strong cation exchange chromatography. Using the separation selectivity, we successfully enrich the protein N-terminal peptides, applying to N-terminomics.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (17 results)

All 2018 2017 2016 Other

All Int'l Joint Research (1 results) Journal Article (1 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (15 results) (of which Int'l Joint Research: 4 results,  Invited: 4 results)

  • [Int'l Joint Research] ベルリン工科大学(ドイツ)

    • Related Report
      2017 Annual Research Report
  • [Journal Article] Peptide Retention in Hydrophilic Strong Anion Exchange Chromatography Is Driven by Charged and Aromatic Residues2018

    • Author(s)
      Giese Sven H.、Ishihama Yasushi、Rappsilber Juri
    • Journal Title

      Analytical Chemistry

      Volume: 90 Issue: 7 Pages: 4635-4640

    • DOI

      10.1021/acs.analchem.7b05157

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] プロテオミクス試料調製用タンパク質固定化StageTipの開発2018

    • Author(s)
      西田紘士・若林真樹・杉山直幸・石濱泰
    • Organizer
      日本薬学会第138年会
    • Related Report
      2017 Annual Research Report
  • [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] Peptide Chromatographic Approaches for Illuminating Human Proteome2018

    • Author(s)
      Y. Ishihama
    • Organizer
      18th KHUPO Annual International Proteomics Conference
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] One-step Enrichment Method for Protein N-Terminome2017

    • Author(s)
      Chih-Hsiang Chang, Eito Yamamoto, Hsin-Yi Chang , Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      AOMSC 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] N-terminal Arg/Lys Proteases for Shotgun Proteomics2017

    • Author(s)
      Chih-Hsiang Chang, 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] タンパク質末端大規模解析に向けた新規タンパク質末端ペプチド濃縮法の開発2017

    • Author(s)
      山本 英人、張 智翔、木村 迪子、杉山 直幸、石濱 泰
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
  • [Presentation] ポリマー被覆モノリス型シリカを用いた逆相系キャピラリーカラムの開発2017

    • Author(s)
      舟橋充央、阿知波弘憲、Hsin-Yi Chang、 杉山直幸、 石濱泰
    • Organizer
      第24回クロマトグラフィーシンポジウム
    • Related Report
      2017 Annual Research Report
  • [Presentation] メートル長C18モノリス型シリカキャピラリーカラムと 緩勾配溶出を組み合わせたnanoLC/MS/MSによる タンパク質同定システムの最適化2017

    • Author(s)
      舟橋充央、阿知波弘憲、山本有希子、Hsin-Yi Chang、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] メートル長HILICモノリス型シリカカラムを用いた高分離能nanoLC/MS/MSプロテオミクス2016

    • Author(s)
      阿知波 弘憲, 鎌倉 健雄, 市原 駿, 若林 真樹, 杉山 直幸, 石濱 泰
    • Organizer
      第27回クロマトグラフィー科学会議
    • Place of Presentation
      慶應義塾大学芝共立キャンパス(東京都港区)
    • Year and Date
      2016-11-16
    • Related Report
      2016 Research-status Report
  • [Presentation] ヒトプロテオーム解明に向けたプロテオーム 解析法の開発2016

    • Author(s)
      石濱 泰
    • Organizer
      第36回キャピラリー電気泳動シンポジウム
    • Place of Presentation
      徳島大学常三島キャンパス(徳島市)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] 腸内細菌叢メタプロテオーム解析プラットフォームの開発2016

    • Author(s)
      伊藤麻里子、若林真樹、杉山直幸、園村和弘、松田文彦、石濱泰
    • Organizer
      第89回日本生化学会大会
    • Place of Presentation
      仙台国際センター(宮城県仙台市)
    • Year and Date
      2016-09-25
    • Related Report
      2016 Research-status Report
  • [Presentation] Acetyl proteomics using a novel enrichment strategy for lysine acetylated peptides2016

    • Author(s)
      Kazuya Tsumagari, Takayuki Hashimoto, Masaki Wakabayashi, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      HUPO2016
    • Place of Presentation
      Taipei, Taiwan
    • Year and Date
      2016-09-18
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] High sensitivity and high throughput phosphoproteomics with micro-scale TMT labeling2016

    • Author(s)
      Kosuke Ogata, Chia-Feng Tsai, Masaki Wakabayashi, Naoyuki Sugiyama, Yasushi Ishihama
    • Organizer
      21st International Mass Spectrometry Conference
    • Place of Presentation
      Toronto, Canada
    • Year and Date
      2016-08-20
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] DIAスライスモードを用いたLC/MS/MSによる大規模定量プロテオミクス2016

    • Author(s)
      阿知波弘憲、若林真樹、杉山直幸、石濱泰
    • Organizer
      日本プロテオーム学会2016年大会
    • Place of Presentation
      北里大学薬学部(東京都港区)
    • Year and Date
      2016-07-28
    • Related Report
      2016 Research-status Report
  • [Presentation] プロテオーム間引き解析とSWATH2016

    • Author(s)
      石濱 泰
    • Organizer
      第64回質量分析総合討論会
    • Place of Presentation
      ホテル阪急エキスポパーク(大阪府吹田市)
    • Year and Date
      2016-05-18
    • Related Report
      2016 Research-status Report
    • Invited

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Published: 2016-04-21   Modified: 2019-03-29  

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