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Development of a method for protein labeling by the use of intramolecular thioester-forming unit

Research Project

Project/Area Number 24550187
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Chemistry related to living body
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) HIRAKI Yuji  京都大学, 再生医科学研究所, 教授 (40144498)
Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords生体分子 / ペプチド / タンパク質 / 受容体 / チオエステル / 部位特異的修飾
Outline of Final Research Achievements

We had found that a peptide containing a sequence, Cys-Pro ester (CPE), at the C terminus is spontaneously transformed into a peptide thioester, which can be used for the peptide ligation. In this research, we aimed to develop a method for the site-specific labeling of proteins.
At first, we made modification of the original CPE structure for the use of protein labeling, in which the ligand for protein binding was retained after thioester formation. Then, a peptide with partial sequence of parathyroid hormone (PTH) containing the modified CPE structure and a fluorescence group was synthesized. When the cell expressing a PTH receptor was treated with the peptide, synthesized, the fluorescence was observed at the same position with the receptor, and we were not able to confirm the covalent bonding between the fluorescence group and the receptor. On the other hand, we succeeded in the total chemical synthesis of proteins using the newly developed modified CPE structure.

Report

(4 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • 2012 Research-status Report
  • Research Products

    (22 results)

All 2015 2014 2013 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Acknowledgement Compliant: 1 results) Presentation (14 results) (of which Invited: 3 results) Remarks (1 results)

  • [Journal Article] Phenacyl Group as a Protecting Group for Thiol in the Use of Recombinant Peptides as Building Blocks for the Peptide Ligation Strategy2015

    • Author(s)
      T. Kawakami, H. Hojo
    • Journal Title

      Peptide Science 2014

      Volume: - Pages: 117-118

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Peptide Thioester Formation via an Intramolecular N toS Acyl Shift for Peptide Ligation2015

    • Author(s)
      T. Kawakami
    • Journal Title

      Topics in Current Chemistry

      Volume: 362 Pages: 107-136

    • DOI

      10.1007/128_2014_575

    • ISBN
      9783319191850, 9783319191867
    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Synthesis of Histone Proteins by CPE Ligation using a Recombinant Peptide as the C-Terminal Building Block2015

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

      J. Biochem.

      Volume: 未定

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Total Chemical Synthesis and Semi-synthesis of Histone H3 and H4 by the Use of Cys-Pro Ester Auto-activating Unit for the Thioester2014

    • Author(s)
      T. Kawakami, Y. Akai, R. Yoshikawa, I. Suetake, S. Tajima, S. Aimoto
    • Journal Title

      Peptide Science 2013

      Volume: - Pages: 47-48

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Enhancement in the Rate of Conversion of Peptide Cys-Pro esters to Peptide Thioesters by Structural Modification2014

    • Author(s)
      T. Kawakami, A. Kamauchi, E. Harada, S. Aimoto
    • Journal Title

      Tetrahedron Lett.

      Volume: 55 Issue: 1 Pages: 79-81

    • DOI

      10.1016/j.tetlet.2013.10.121

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Sequential Peptide Ligation by Combining the Cys-Pro Ester (CPE) and Thioester Methods and its Application to the Synthesis of Histone H3 Containing a Trimethyl Lysine Residue2013

    • Author(s)
      T. Kawakami, Y. Akai, H. Fujimoto, C. Kita, Y. Aoki, T. Konishi, M. Waseda, L. Takemura, S. Aimoto
    • Journal Title

      Bull. Chem. Soc. Jpn

      Volume: 86 Issue: 6 Pages: 690-697

    • DOI

      10.1246/bcsj.20130026

    • NAID

      10031176002

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Sequential Peptide Ligation by the Combination of Cys-Pro Ester and Thioester Methods: An Application to a Synthesis of Histone H32013

    • Author(s)
      T. Kawakami, Y. Akai, H. Fujimoto, C. Kita, L. Takemura, Y. Aoki, M. Waseda, S. Aimoto
    • Journal Title

      Peptide Science 2012

      Volume: - Pages: 77-78

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Presentation] ライゲーション法において発現ペプチドを合成ブロックとして用いるためのフェナシル基のチオール保護基としての可能性2015

    • Author(s)
      川上 徹,北條裕信
    • Organizer
      日本化学会第95春季年会
    • Place of Presentation
      千葉県船橋市 日本大学
    • Year and Date
      2015-03-26
    • Related Report
      2014 Annual Research Report
  • [Presentation] Methods for Peptide Thioester Preparation2015

    • Author(s)
      Toru Kawakami
    • Organizer
      7th Annual AsiaTIDE
    • Place of Presentation
      大阪府大阪市 ハイアットリージェンシー大阪
    • Year and Date
      2015-03-04
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 発現ペプチドをライゲーション法の合成ブロックとして用いるためのフェナシル基のチオール保護基としての可能性2014

    • Author(s)
      川上 徹,北條裕信
    • Organizer
      第51回ペプチド討論会
    • Place of Presentation
      徳島県徳島市 徳島大学
    • Year and Date
      2014-10-23
    • Related Report
      2014 Annual Research Report
  • [Presentation] Synthesis of modified histones by the use of CPE chemistry2014

    • Author(s)
      Toru Kawakami
    • Organizer
      The 15th Akabori Conference
    • Place of Presentation
      ドイツ ボッパード  ベストウエスティン
    • Year and Date
      2014-09-09
    • Related Report
      2014 Annual Research Report
  • [Presentation] Synthesis of Histone by the Use of Cys-Pro Ester (CPE) Ligation2014

    • Author(s)
      Toru Kawakami, Ryo Yoshikawa, Yuki Fujiyoshi, Hironobu Hojo, Shoji Tajima, Isao Suetake
    • Organizer
      33th European Peptide Symposium
    • Place of Presentation
      ブルガリア ソフィア 国立文化宮殿
    • Year and Date
      2014-09-04
    • Related Report
      2014 Annual Research Report
  • [Presentation] ケミカルライゲーションによる修飾ヒストン合成法の開発:H4K20meの合成とテトラソームの再構成2013

    • Author(s)
      川上 徹,吉川 諒,相本三郎,田嶋正二,末武 勲
    • Organizer
      エピジェネティクス研究会 第7回年会
    • Place of Presentation
      奈良県新公会堂,奈良
    • Related Report
      2013 Research-status Report
  • [Presentation] Protein synthesis by chemical ligation base on the use of a peptide-Cys-Pro ester as a peptide thioester precursor2013

    • Author(s)
      T. Kawakami, S. Aimoto
    • Organizer
      International Scientific Conference on “Chemistry - Opportunity and Development” 35th Anniversary of the Institute of Chemistry
    • Place of Presentation
      The Institute of Chemistry, Vietnam Academy of Science and Technology, Hanoi, Vietnam
    • Related Report
      2013 Research-status Report
    • Invited
  • [Presentation] Rate Enhancement of Conversion of Peptide Cys-Pro ester to Peptide Thioester by the Structural Modifications2013

    • Author(s)
      T. Kawakami, A. Kamauchi, E. Harada, S. Aimoto
    • Organizer
      4th Modern Solid Phase Peptide Synthesis & Its Applications Symposium
    • Place of Presentation
      Hotel Maiko Villa Kobe, Kobe
    • Related Report
      2013 Research-status Report
  • [Presentation] Total Chemical Synthesis and Semi-synthesis of Histone H3 and H4 by the Use of Cys-Pro Ester Auto-activating Unit for the Thioester Precursor2013

    • Author(s)
      T. Kawakami, Y. Akai, R. Yoshikawa, I. Suetake, S. Tajima, S. Aimoto
    • Organizer
      5th Asia-Pacific International Peptide Symposium
    • Place of Presentation
      Hotel Hankyu Expopark, Suita
    • Related Report
      2013 Research-status Report
  • [Presentation] Synthesis of Histone H3 by Chemical Ligation2013

    • Author(s)
      T. Kawakami
    • Organizer
      The 8th International Conference on Cutting-Edge Organic Chemistry in Asia (ICCEOCA-8)
    • Place of Presentation
      Grand Cube, Osaka
    • Related Report
      2013 Research-status Report
  • [Presentation] Chemical Protein Synthesis by the Ligation Based on the Use of an N to S Acyl Shift Reaction2013

    • Author(s)
      T. Kawakami
    • Organizer
      17th Korean Peptide and Protein Symposium
    • Place of Presentation
      Seoul National University, Korea
    • Related Report
      2013 Research-status Report
    • Invited
  • [Presentation] Peptide thioester formation at a Cys residue via an N-S acyl shift reaction

    • Author(s)
      Toru Kawakami, Akitaka Kamauchi, Saburo Aimoto
    • Organizer
      The 14th Akabori Conference
    • Place of Presentation
      北海道ニセコ町民センター
    • Related Report
      2012 Research-status Report
  • [Presentation] Cys-Proエステル法とチオエステル法の連続ライゲーション:ヒストンH3の合成

    • Author(s)
      川上 徹,赤井優一,藤本久雄,喜多千恵子,竹村梨沙,青木裕子,早稲田真澄,相本三郎
    • Organizer
      第49回ペプチド討論会
    • Place of Presentation
      鹿児島県民交流センター
    • Related Report
      2012 Research-status Report
  • [Presentation] 自発的活性化ユニットCys-Proエステルを用いる連続ライゲーションによるヒストンH3の合成

    • Author(s)
      川上 徹・赤井優一・藤本久雄・喜多千恵子・竹村梨沙・青木優子・早稲田真澄・相本三郎
    • Organizer
      日本化学会第93春季年会
    • Place of Presentation
      滋賀県草津市 立命館大学びわこ・くさつキャンパス
    • Related Report
      2012 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
      2014 Annual Research Report

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Published: 2013-05-31   Modified: 2019-07-29  

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