High sensitive detection of ubiquitination reaction in urine

• Project/Area Number: 23700588
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Medical systems
• Research Institution: Himeji Dokkyo University
• Principal Investigator: MIYAMOTO Kazuhide 姫路獨協大学, 薬学部, 准教授 (10415317)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2011: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 検査 / 診断システム ユビキチン化 / ユビキチン化

Research Abstract

Protein ubiquitination is undertaken through an enzymatic cascade comprising ubiquitin-activating (E1), ubiquitin-conjugating (E2), and ubiquitin-ligating (E3) enzymes. This ubiquitination reaction is related to several diseases such as bladder cancer, prostate cancer, kidney cancer, leukemia. The high sensitive detection of the degree of the ubiquitination leads to the diagnosis of these diseases. In this study, (1) the artificial E3 was designed for the detection of the ubiquitination without a substrate and a tag (2) the micro bioactivity analyzer (AMIS-101) had a high sensitivity for detecting the degree of the ubiquitination of the artificial E3 in the solution including the purified ubiquitin, E1, E2. (3) in human acute myelogenous leukemia cells with treatment of the cancer drugs, the detection system of the ubiquitination of the artificial E3 using AMIS-101 made it possible to the observation of the small amount of the E2 enzyme activity.

Research Products

• [Journal Article] Oligopeptides derived from autophosphorylation sites of EGF receptor suppress EGF-stimulated responses in human lung carcinoma A549 cells. (2013)
  • Author(s): Kuroda Y, Kato-Kogoe N, Tasaki E, Murata M, Ueda K, Abe M, Miyamoto K, Nakase I, Futaki S, Tohyama Y, Hirose M
  • Journal Title: Eur J Pharmacol Volume: 698 Issue: 1-3 Pages: 87-94
  • DOI: 10.1016/j.ejphar.2012.10.007
  • Peer Reviewed
• [Journal Article] T アレルギー性皮膚炎の治療薬-シコニンの効果 (2013)
  • Author(s): Watanabe A, Hichiri H, Takenokuchi M, Miyamoto K, Kadoyama K, Taniguchi
  • Journal Title: Small Animal Dermatology Volume: 9 Pages: 60-65
  • Peer Reviewed
• [Journal Article] アレルギー性皮膚炎の治療薬－シコニンの効果― (2013)
  • Author(s): Watanabe A, Hichiri H, Takenokuchi M, Miyamoto K, Kadoyama K, Taniguchi T
  • Journal Title: Small Animal Dermatology Volume: 9 Pages: 60-65
  • Peer Reviewed
• [Journal Article] Ubiquitination of an artificial RING finger without a substrate and a tag (2012)
  • Author(s): K. Miyamoto
  • Journal Title: Journal of Peptide Science Volume: 18 Issue: 2 Pages: 135-139
  • DOI: 10.1002/psc.1426
  • Peer Reviewed
• [Journal Article] TogiyaSolution structure of LC4 transmembrane segment of CCR5 (2011)
  • Author(s): K. Miyamoto and K.
  • Journal Title: PLoS One Volume: 6 Issue: 5 Pages: e20452-e20452
  • DOI: 10.1371/journal.pone.0020452
  • Peer Reviewed
• [Presentation] An artificial ubiquitin-ligating (E3) enzyme by the method of alpha helical region substitution (2012)
  • Author(s): Miyamoto K
  • Organizer: 第35回日本分子生物学会
  • Place of Presentation: 福岡国際会議場(九州)
  • Year and Date: 2012-12-11
• [Presentation] An artificial ubiquitin-ligating (E3) enzyme by the method of alpha helical region substitution (2012)
  • Author(s): Miyamoto K
  • Organizer: 第35回日本分子生物学会
  • Place of Presentation: 福岡国際会議場(九州）
• [Presentation] Creation of an artificial RING finger without a substrate and a tag (2011)
  • Author(s): Miyamoto K
  • Organizer: 第48回ペプチド討論会
  • Place of Presentation: 札幌コンベンションセンター(北海道)
  • Year and Date: 2011-09-28
• [Presentation] Creation of an artificial RING finger without a substrate and a tag (2011)
  • Author(s): 宮本和英
  • Organizer: 第48回ペプチド討論会
  • Place of Presentation: 札幌コンベンションセンター（北海道）