Development of new synthetic method of ubiquitinated glycoprotein probe using specific reactivity of selenium

• Project/Area Number: 17H02211
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Chemical biology
• Research Institution: Kochi University
• Principal Investigator: Izumi Masayuki 高知大学, 教育研究部総合科学系複合領域科学部門, 教授 (80332641)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2020: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2019: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2018: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2017: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
• Keywords: 精密化学合成 / ユビキチン / セレン / 連結反応 / 糖タンパク質 / 分解 / 脱セレノ化 / 分子プローブ / 変性糖タンパク質

Outline of Final Research Achievements

The development of ubiquitinated glycoprotein probes is important for the study of the degradation of aberrant glycoproteins by ubiquitin-proteasome system in the cell. Efficient chemical ubiquitination is required to synthesize ubiquitinated glycoprotein probes. We developed a novel δ-selenolysine-mediated ligation reaction with ubiquitin-α-thioester that does not affect cysteine thiols. Fmoc-δ-seleno- L-lysine and D-lysine derivatives were synthesized from δ-hydroxy-DL-lysine via enzymatic optical resolution. Ubiquitin peptide containing δ-seleno-L-lysine was synthesized by solid-phase peptide synthesis, and one-pot ligation-deselenization was achieved with ubiquitin peptide-α-thioester to yield K48-linked diubiquitin peptide. This new δ-selenolysine-mediated ligation-deselenization strategy will be useful in the synthesis of ubiquitinated glycoprotein probes.

Academic Significance and Societal Importance of the Research Achievements

糖タンパク質の分解経路の異常は、細胞の恒常性の維持に影響を与え神経変性疾患などの原因ともなる。現在盛んに開発されているタンパク質のユビキチン化反応に、本研究で開発されたδ-セレノリシン残基とユビキチン-α-チオエステルとの化学選択的連結-脱セレノ化反応が新たに加わることで、ユビキチン化糖タンパク質などより複雑な構造を持ったプローブの合成が効率的になり、ユビキチンバイオロジーの発展がさらに加速される。また、合成したプローブを用いた変性糖タンパク質分解経路の解明を通して基礎生物医学的な応用も期待される。

Research Products

• [Journal Article] Chemical Synthesis of Ubiquitinated High-Mannose-Type N-Glycoprotein CCL1 in Different Folding States (2020)
  • Author(s): Izumi Masayuki、Araki Hiroyuki、Tominaga Mamiko、Okamoto Ryo、Kajihara Yasuhiro
  • Journal Title: The Journal of Organic Chemistry Volume: 85 Issue: 24 Pages: 16024-16034
  • DOI: 10.1021/acs.joc.0c01766
  • Peer Reviewed
• [Presentation] Chemical synthesis of diubiquitin using delta-selenolysine-mediated ligation (2021)
  • Author(s): Akiyama, Tatsunari; Tanaka, Yusuke; Okamoto, Ryo; Kajihara, Yasuhiro; Izumi, Masayuki
  • Organizer: Pacifichem2021
  • Int'l Joint Research
• [Presentation] Fmoc-δ-セレノリジン誘導体の合成法の改良 (2019)
  • Author(s): 秋山龍成、和泉雅之
  • Organizer: 高知化学シンポジウム2019
• [Presentation] ユビキチン化糖鎖化インターロイキン８の合成研究 (2019)
  • Author(s): ジョンソンエマリー、和泉雅之
  • Organizer: 高知化学シンポジウム2019
• [Presentation] ペプチド固相合成に利用できる δ 位に置換基を有する D リジン誘 導体の合成 (2019)
  • Author(s): 秋山 龍成、田中 勇祐、岡本 亮、梶原 康宏、和泉 雅之
  • Organizer: 日本化学会第９９春季年会
• [Presentation] Selenoisopeptide Chemical Ligation toward Chemical Synthesis of Homogeneous Ubiquitinated Glycoprotein Containing Disufide Bond (2018)
  • Author(s): Izumi, M., Tanaka, Y., Araki, H., Maki, Y., Okamoto, R., Kajihara, Y.
  • Organizer: 29th International Carbohydrate Symposium
  • Int'l Joint Research
• [Presentation] δ-セレノリジン誘導体を用いたユビキチン化糖タンパク質の合成研究 (2018)
  • Author(s): 田中勇祐、真木勇太、岡本亮、梶原康宏、和泉雅之
  • Organizer: 日本化学会第98春季年会