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Synthesis of histone protein for the post-translationally modified protein library

Research Project

Project/Area Number 15K05565
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Bio-related chemistry
Research InstitutionOsaka University

Principal Investigator

Kawakami Toru  大阪大学, たんぱく質研究所, 准教授 (70273711)

Co-Investigator(Renkei-kenkyūsha) SUETAKE Isao  甲子園大学, 栄養学部, 教授 (80304054)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsヒストン / 翻訳後修飾 / ペプチドライゲーション / イソペプチド / チイラン / ユビキチン / DNAメチル化
Outline of Final Research Achievements

The site specific modification of proteins such as post-translational modification (PTM) is important for studies of protein functions. The PTMs of histones play an important role in the epigenetic regulation. In this research we aimed to develop the methods for preparation of the site specifically modified histones by using the CPE ligation method.
At first, a semi-synthetic method was developed, in which recombinant C-terminal peptides were ligated with the N-terminal site specifically modified CPE peptides, followed by desulfurized to produce trimethyl lysine containing histones H3 and H4. We also developed a convenient method for preparation of isopeptide mimetics by using a thiirane linker. Using this method, ubiquitinated histone H3 peptide and full length H3 were prepared, and these peptides were used for their structural and functional studies. In addition, in order to develop more convenient synthetic method for histones preliminary result of solid phase peptide ligation.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (19 results)

All 2018 2017 2016 2015 Other

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 3 results,  Acknowledgement Compliant: 2 results) Presentation (12 results) (of which Int'l Joint Research: 3 results,  Invited: 1 results) Remarks (1 results)

  • [Journal Article] 1.Synthesis of ubiquitylated histone H3 using a thiirane linker for chemical ligation2017

    • Author(s)
      T. Kawakami, Y. Mishima, H. Hojo, I Suetake
    • Journal Title

      J. Pept. Sci.

      Volume: 印刷中 Issue: 7-8 Pages: 532-538

    • DOI

      10.1002/psc.2976

    • Related Report
      2017 Annual Research Report 2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] RFTS-dependent negative regulation of Dnmt1 by nucleosome structure and histone tails.2017

    • Author(s)
      Mishima Y, Brueckner L, Takahashi S, Kawakami T, Arita K, Oka S, Otani J, Hojo H, Shirakawa M, Shinohara A, Watanabe M, Suetake I
    • Journal Title

      FEBS J.

      Volume: 284 Issue: 20 Pages: 3455-3469

    • DOI

      10.1111/febs.14205

    • NAID

      120006537075

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Structure of the Dnmt1 reader module complexed with a unique two-mono-ubiquitin mark on histone H3 reveals the basis for DNA methylation maintenance.2017

    • Author(s)
      Ishiyama S, Nishiyama A, Saeki Y, Moritsugu K Morimoto D, Yamaguchi L, Arai N, Matsumura R, Kawakami T, Mishima Y, Hojo H, Shimamura S, Ishikawa F, Tajima S, Tanaka K, Ariyoshi M, Shirakawa M, Ikeguchi M, Kidera A, Suetake I, Arita K, Nakanishi M
    • Journal Title

      Mol Cell

      Volume: 68 Issue: 2 Pages: 350-360

    • DOI

      10.1016/j.molcel.2017.09.037

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] One-pot native chemical ligation by combination of two orthogonal thioester precursors2017

    • Author(s)
      Y. Asahina, Toru Kawakami and Hironobu Hojo
    • Journal Title

      Chem. Commun

      Volume: 53 Issue: 13 Pages: 2114-2117

    • DOI

      10.1039/c6cc10243c

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] A thiirane linker for isopeptide mimetics by peptide ligation2016

    • Author(s)
      T. Kawakami, Y. Mishima, M. Kinoshita, Y.-H. Lee, I. Suetake
    • Journal Title

      Tetrahedron Letters

      Volume: 57 Issue: 19 Pages: 2112-2115

    • DOI

      10.1016/j.tetlet.2016.04.006

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Synthesis of modified histone proteins by CPE ligation using a recombinant peptide2016

    • Author(s)
      T. Kawakami, R. Yoshikawa, Y. Fujiyoshi, Y. Mishima, H. Hojo, S. Tajima, I. Suetake
    • Journal Title

      Peptide Science 2015

      Volume: - Pages: 117-118

    • Related Report
      2015 Research-status Report
    • Peer Reviewed
  • [Presentation] チイランリンカーを用いる蛋白質の簡便なユビキチン化2018

    • Author(s)
      川上徹,三島優一,北條裕信,末武勲
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Histone Ubiquitination via an Isopeptide Mimetic Structure by Using a Thiirane Linker2017

    • Author(s)
      川上徹,三島優一,北條裕信,末武勲
    • Organizer
      第54回ペプチド討論会
    • Related Report
      2017 Annual Research Report
  • [Presentation] チイランリンカーを用いるイソペプチド類似構造を介したユビキチン化ヒストンタンパク質の調製2017

    • Author(s)
      川上徹,三島優一,北條裕信,末武勲
    • Organizer
      2017 年度生命科学系学会合同年次大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] ユビキチン化ヒストンは、維持型メチル化酵素Dnmt1のメチル化維持活性を促進する2017

    • Author(s)
      三島優一,川上徹,北條裕信,有田恭平,末武勲
    • Organizer
      2017 年度生命科学系学会合同年次大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Ubiquitination of Histone Peptides via Isopeptide Mimetics by Using a Thiirane Linker2016

    • Author(s)
      T. Kawakami, Y. Mishima, M. Kinoshita, Y.-H. Lee, I. Suetake
    • Organizer
      第53回ペプチド討論会
    • Place of Presentation
      京都テルサ(京都府京都市)
    • Year and Date
      2016-10-16
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] チイランリンカーを用いるイソペプチド類似構造を介したヒストンペプチドのユビキチン化2016

    • Author(s)
      川上 徹,三島優一,木下 岬,李 映昊,末武 勲
    • 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] チイランリンカーを用いたペプチドライゲーション法によるイソペプチド類似体の合成2016

    • Author(s)
      川上 徹,三島優一,木下 岬,李 映昊,末武 勲
    • Organizer
      第10回バイオ関連化学シンポジウム
    • Place of Presentation
      石川県立音楽堂(石川県金沢市)
    • Year and Date
      2016-09-07
    • Related Report
      2016 Research-status Report
  • [Presentation] Synthesis of Modified Histone Proteins by CPE Ligation2016

    • Author(s)
      T. Kawakami
    • Organizer
      The 16th Akabori Conference
    • Place of Presentation
      六甲山ホテル(兵庫県神戸市)
    • Year and Date
      2016-05-24
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] ライゲーション法によるイソペプチド類似構造を介したヒストンペプチドのユビキチン化2016

    • Author(s)
      川上 徹,三島優一,木下 岬,李 映昊,末武 勲
    • Organizer
      エピジェネティクス研究会 第10回年会
    • Place of Presentation
      千里ライフサイエンスセンター(大阪府豊中市)
    • Year and Date
      2016-05-19
    • Related Report
      2016 Research-status Report
  • [Presentation] Synthesis of modified histone proteins by CPE ligation using a recombinant peptide2015

    • Author(s)
      T. Kawakami, R. Yoshikawa, Y. Fujiyoshi, Y. Mishima, H. Hojo, S. Tajima, I. Suetake
    • Organizer
      7th International Peptide Symposium
    • Place of Presentation
      Singapore (Singapore)
    • Year and Date
      2015-12-09
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] ヒストンの化学合成2015

    • Author(s)
      川上 徹
    • Organizer
      BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会)
    • Place of Presentation
      神戸国際会議場(兵庫県神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] Synthesis of modified histone proteins by CPE ligation using a recombinant peptide2015

    • Author(s)
      T. Kawakami, R. Yoshikawa, Y. Fujiyoshi, Y. Mishima, H. Hojo, S. Tajima, I. Suetake
    • Organizer
      第52回ペプチド討論会
    • Place of Presentation
      平塚中央公民館(神奈川県平塚市)
    • Year and Date
      2015-11-16
    • Related Report
      2015 Research-status Report
  • [Remarks] 出版物(川上徹) | 蛋白質有機化学研究室

    • URL

      http://www.protein.osaka-u.ac.jp/organic/publication/kawa.html

    • Related Report
      2015 Research-status Report

URL: 

Published: 2015-04-16   Modified: 2019-03-29  

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