Development of profiling method for O-glycopeptides

• Project/Area Number: 17K15097
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Japanese Foundation for Cancer Research
• Principal Investigator: HAGA Yoshimi 公益財団法人がん研究会, がんプレシジョン医療研究センター がんオーダーメイド医療開発プロジェクト, 研究員 (40525789)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2017: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 糖鎖 / 質量分析 / グライコプロテオミクス / 癌 / プロテオーム

Outline of Final Research Achievements

To identify cancer-specific O-linked glycosylated proteins comprehensively, we developed a profiling technology that quantify and profile O-linked glycosylation sites in a very small amount of biological intact samples. We achieved high enrichment efficiency of O-glycopeptides and was able to identify plural unreported O-linked glycosylation sites on a model protein which was known to be multiply O-glycosylated. Furthermore, by the analysis using a cancer cell extract, we confirmed that cancer-related molecules such as mucins and NF-κB are certainly O-glycosylated.

Academic Significance and Societal Importance of the Research Achievements

糖鎖のうちN型糖鎖は付加部位のコンセンサス配列や基幹構造と呼ばれる共通の基礎構造があり、全N型糖鎖を遊離可能な酵素PNGase-Fも知られているため比較的研究がよく進んでいる。しかし、O型糖鎖はこれらの規則、ツールが全て無く、従来法や最新の質量分析計を駆使しても単一タンパク質のO型糖鎖付加部位を決定することさえ困難な状況であった。本開発技術は、あらゆる生体サンプル中から網羅的にO型糖鎖付加部位の同定と付加頻度の定量化を一度の質量分析で同時に可能とする世界初の分析手法である。

Research Products

• [Journal Article] Identification of Multisialylated LacdiNAc Structures as Highly Prostate Cancer Specific Glycan Signatures on PSA. (2019)
  • Author(s): Haga, Y., Uemura, M., Baba, S., Inamura, K., Takeuchi, K., Nonomura, N., and Ueda, K.
  • Journal Title: Analytical chemistry Volume: 91 Issue: 3 Pages: 2247-2247
  • DOI: 10.1021/acs.analchem.8b04829
  • Peer Reviewed
• [Presentation] 前立腺癌特異的PSA糖鎖構造の同定による高特異度前立腺癌マーカーPSA G-indexの開発 (2018)
  • Author(s): 芳賀淑美、植村元秀、稲村健太郎、竹内賢吾、野々村祝夫、植田幸嗣
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] Rapid profiling of prostate cancer-specific PSA glycoforms as a specificity-enhanced secondary biomarker (2018)
  • Author(s): Yoshimi Haga, Motohide Uemura, Kentaro Inamura, Kengo Takeuchi, Norio Nonomura and Koji Ueda
  • Organizer: HUPO 2018
  • Int'l Joint Research
• [Presentation] Identification of highly prostate cancer-specific PSA glyco-isoforms (PSA G-index) (2017)
  • Author(s): Yoshimi Haga, Motohide Uemura, Norio Nonomura and Koji Ueda
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] Identification of highly prostate cancer-specific PSA glyco-isoforms (PSA G-index) (2017)
  • Author(s): Yoshimi Haga, Motohide Uemura, Norio Nonomura and Koji Ueda
  • Organizer: International Symposium"Systems Glycobiology and Beyond"
  • Int'l Joint Research