Peptidomics for studying limited proteolysis in pancreatic cancer

• Project/Area Number: 16K09404
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Sasaki Foundation (2018); National Cardiovascular Center Research Institute (2016-2017)
• Principal Investigator: Sasaki Kazuki 公益財団法人佐々木研究所, 附属研究所, 部長(移行) (80260313)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 膵腺癌 / ペプチド / 限定分解 / 質量分析 / タンパク質切断 / ペプチドミクス / 膵臓学

Outline of Final Research Achievements

We used mass spectrometry-based profiling of endogenous peptides released in medium by cultured pancreatic cancer cell lines and immortalized pancreatic duct cell lines in order to identify specific, extracellular proteolytic processing sites of secretory or transmembrane proteins. Sequenced peptides were aligned on each parent precursor to generate proteolytic processing maps. Some known processing sites such as peptide hormone processing or transmembrane protein ectodomain shedding were recapitulated by this peptidomic profiling. In-depth analysis of endogenous peptides would highlight more transmembrane proteins regulated by ectodomain shedding. Accumulated profiling information can be used for identifying transmembrane proteins that could be a potential target for treating pancreatic cancer.

Academic Significance and Societal Importance of the Research Achievements

膜タンパク質のプロセシング部位を特定することは、新規生理活性ペプチドの生成部位の手がかりを提供するだけではなく、抗体医薬の設計に必要な基盤情報の提供にも役立つ。従来は部位の特定は、個別的な生化学・分子生物学的な研究で実施されてきた。しかし、本研究では、質量分析法の進歩で可能となったペプチドミクスの手法を活用することで、膜タンパク質のプロセシング部位をトランスフェクションなどを介することなく、そのままの形で特定できるようになったことが大きな進歩である。深達度の高い解析で、治療の標的となる分子の特定が期待される。

Research Products

• [Journal Article] Isolation of Endogenous Peptides from Cultured Cell Conditioned Media for Mass Spectrometry (2018)
  • Author(s): Sasaki Kazuki、Tsuchiya Takashi、Osaki Tsukasa
  • Journal Title: Methods in Molecular Biology Volume: 1719 Pages: 51-58
  • DOI: 10.1007/978-1-4939-7537-2_3
  • ISBN: 9781493975365, 9781493975372
  • Peer Reviewed
• [Journal Article] Endogenous peptide profile for elucidating biosynthetic processing of the ghrelin precursor (2017)
  • Author(s): Tsuchiya Takashi、Iwakura Hiroshi、Minamino Naoto、Kangawa Kenji、Sasaki Kazuki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 490 Issue: 4 Pages: 1142-1146
  • DOI: 10.1016/j.bbrc.2017.06.155
  • Peer Reviewed
• [Journal Article] ペプチドミクスを活用した生理活性ペプチドの発見 (2017)
  • Author(s): 佐々木一樹
  • Journal Title: 生化学 Volume: 89 Pages: 894898-894898
  • NAID: 40021439312
• [Journal Article] NERP-2 regulates gastric acid secretion and gastric emptying via the orexin pathway. (2017)
  • Author(s): Cherl Namkoong, Koji Toshinai, Zaved Waise, Hideyuki Sakoda, Kazuki Sasaki, Yoichi Ueta, Min-Seon Kim, Naoto Minamino, Masamitsu Nakazato.
  • Journal Title: Biochem Biophys Res Commun Volume: 485 Issue: 2 Pages: 409-413
  • DOI: 10.1016/j.bbrc.2017.02.064
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characterization of gonadotrope secretoproteome identifies neurosecretory protein VGF-derived peptide suppression of follicle-stimulating hormone gene expression (2016)
  • Author(s): Choi SG, Wang Q, Jia J, Chikina M, Pincas H, Dolios G, Sasaki K, Wang R, Minamino N, Salton SR, Sealfon SC
  • Journal Title: Journal of Biological Chemistry Volume: 291 Pages: 2132221334-2132221334
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Endogenous Peptide Profiling for Studying Limited Proteolysis (2017)
  • Author(s): Kazuki Sasaki
  • Organizer: PepCon2017
  • Place of Presentation: 福岡
  • Year and Date: 2017-03-23
  • Int'l Joint Research
• [Remarks] 公益財団法人佐々木研究所附属佐々木研究所ペプチドミクス研究部
  • URL: http://www.sasaki-institute.org/peptidomics/