Method for efficient synthesis of chemically functionalization proteins and application for biology research

• Project/Area Number: 19K15705
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37020:Chemistry and chemical methodology of biomolecules-related
• Research Institution: Nagoya University
• Principal Investigator: Oishi Shunsuke 名古屋大学, トランスフォーマティブ生命分子研究所, 特任助教 (80707795)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: タンパク質化学 / ペプチド化学 / ケミカルバイオロジー / 有機合成化学

Outline of Research at the Start

タンパク質の化学修飾は基礎生物学や創薬研究を支える重要な技術となっている。しかし 構造均一かつ精密に修飾されたタンパク質の誘導体群を迅速に得る方法は未開拓である。申請者らは、この問題を解決するため、合成の終盤に誘導化を行うLate-Stage Functionalization を提案する。すなわち化学合成による精密な改変タンパク質の合成とバイオコンジュゲーション化学を組み合わせた戦略である。誘導化に用いる反応剤を変更することにより、簡便に様々な修飾タンパク質を得ることができる。これにより、ケミカルバイオロジー研究や基礎生物学への貢献が期待できる。

Outline of Final Research Achievements

We developed a Late-Stage Functionalization method for chemically synthesized proteins, aiming to systematically and exhaustively synthesize various uniformly structured modified proteins. The distinguishing feature of this study is the ability to obtain modified proteins with completely uniform structures by chemically synthesizing proteins and controlling the reactivity of side chain functional groups.
We succeeded in fluorescently modifying a plant cysteine-rich protein, and conducted bioactivity assays and bioimaging experiments.

Academic Significance and Societal Importance of the Research Achievements

タンパク質の合成では、高純度のタンパク質が得られる反面、しばしば面倒で時間がかかるプロセスである。特に、化学修飾を組み込む場合は、合成初期段階で修飾構造を導入する必要があるため、全合成プロセスを繰り返すことが必要となる。本研究ではLate-Stage Functionalization法により、迅速に修飾タンパク質を化学合成する方法を開発した。

Research Products

• [Int'l Joint Research] ETH Zurich(スイス)
• [Int'l Joint Research] The University of Texas at Austin(米国)
• [Int'l Joint Research] ETH Zurich(スイス)
• [Journal Article] Chemical synthesis of <i>Torenia</i> plant pollen tube attractant proteins by KAHA ligation (2022)
  • Author(s): Kumarswamyreddy Nandarapu、Reddy Damodara N.、Robkis D. Miklos、Kamiya Nao、Tsukamoto Ryoko、Kanaoka Masahiro M.、Higashiyama Tetsuya、Oishi Shunsuke、Bode Jeffrey W.
  • Journal Title: RSC Chemical Biology Volume: 1 Issue: 6 Pages: 1-7
  • DOI: 10.1039/d2cb00039c
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chemical synthesis of the EPF-family of plant cysteine-rich proteins and late-stage dye attachment by chemoselective amide-forming ligations (2022)
  • Author(s): Kumarswamyreddy Nandarapu、Nakagawa Ayami、Endo Hitoshi、Shimotohno Akie、Torii Keiko U.、Bode Jeffrey W.、Oishi Shunsuke
  • Journal Title: RSC Chemical Biology Volume: 3 Issue: 12 Pages: 1422-1431
  • DOI: 10.1039/d2cb00155a
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulation of ammonium acquisition and use in Oryza longistaminata ramets under nitrogen source heterogeneity (2022)
  • Author(s): Misato Kawai, Ryo Tabata, Miwa Ohashi, Haruno Honda, Takehiro Kamiya, Mikiko Kojima, Yumiko Takebayashi, Shunsuke Oishi, Satoru Okamoto, Takushi Hachiya, Hitoshi Sakakibara
  • Journal Title: Plant Physiology Volume: 188 Issue: 4 Pages: 2364-2376
  • DOI: 10.1093/plphys/kiac025
  • Peer Reviewed / Open Access
• [Journal Article] Acceptor-Controlled Transfer Dehydration of Amides to Nitriles (2019)
  • Author(s): Okabe Hiroyuki、Naraoka Asuka、Isogawa Takahiro、Oishi Shunsuke、Naka Hiroshi
  • Journal Title: Organic Letters Volume: 21 Issue: 12 Pages: 4767-4770
  • DOI: 10.1021/acs.orglett.9b01657
  • Peer Reviewed
• [Journal Article] Chemical Synthesis of Atomically Tailored SUMO E2 Conjugating Enzymes for the Formation of Covalently Linked SUMO?E2?E3 Ligase Ternary Complexes (2019)
  • Author(s): Zhang Yinfeng、Hirota Tsuyoshi、Kuwata Keiko、Oishi Shunsuke、Gramani Subramanian G.、Bode Jeffrey W.
  • Journal Title: Journal of the American Chemical Society Volume: 141 Issue: 37 Pages: 14742-14751
  • DOI: 10.1021/jacs.9b06820
  • Peer Reviewed / Int'l Joint Research