Systematic discovery of N-end-rule substrate proteins

• Project/Area Number: 22685017
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Chemistry related to living body
• Research Institution: The University of Electro-Communications (2011-2013); Okayama University (2010)
• Principal Investigator: TAKI Masumi 電気通信大学, 情報理工学(系)研究科, 准教授 (70362952)
• Project Period (FY): 2010-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥24,570,000 (Direct Cost: ¥18,900,000、Indirect Cost: ¥5,670,000); Fiscal Year 2013: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2012: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2011: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2010: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
• Keywords: N末端ルール / NEXT-A反応 / L/F-転移酵素 / 不安定化蛋白質 / tRNA / アミノアシルtRNA合成酵素（ARS) / アミノアシルtRNA合成酵素(ARS) / ユビキチン化酵素(Ubr1) / Huisgen付加反応 / ユビキチン化酵素(Ubrl)

Research Abstract

i) A mutant yeast strain that defects protein degradation system (i.e. ubiquitin ligase) survives when a destabilizing protein is accumulated inside the cell. At that time, the cell overexpresses Obr1 protein to overcome the stressed condition. ii) Kinetic analysis of L/F-transferase with N-end-rule substrate peptides posessing different N-terminal penultimate residues is systematically performed. This analysis suggests that the side chain of the residue affects substrate-binding afffinity towards the transferase. It gives biological insight into the effect of the penultimate amino acid on substrate specificity of natural proteins to be degraded via the N-end rule pathway.

Research Products

• [Journal Article] Construction of a Peptide Expression Vector Library for Phenotypic Screening of Bioactive Peptides in Yeast (2014)
  • Author(s): K. Fukunaga, and M. Taki
  • Journal Title: Peptide Science 2013 Pages: 421-422
  • Peer Reviewed
• [Journal Article] Construction of a Peptide Expression Vector Library for Phenotypic Screening of Bioactive Peptides in Yeast (2014)
  • Author(s): K. Fukunaga* and M. Taki*
  • Journal Title: Peptide Science Volume: 2013 Pages: 421-422
  • Peer Reviewed
• [Journal Article] Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues (2013)
  • Author(s): J. Kawaguchi, K. Maejima, H. Kuroiwa, and M. Taki
  • Journal Title: FEBS Open Bio Volume: 3 Issue: 1 Pages: 252-255
  • DOI: 10.1016/j.fob.2013.06.001
  • Peer Reviewed
• [Journal Article] Fission yeast Ubr1 ubiquitin ligase influences the oxidative stress response via degradation of active Pap1 bZIP transcription factor in the nucleus (2011)
  • Author(s): K. Kitamura, M. Taki, N. Tanaka, I. Yamashita
  • Journal Title: Mol. Microbiol Volume: 80 Issue: 3 Pages: 739-755
  • DOI: 10.1111/j.1365-2958.2011.07605.x
  • Peer Reviewed
• [Journal Article] Synthesis of cyclic peptides using the NEXT-A Reaction (2011)
  • Author(s): T. Hamamoto, M. Sisido, T. Ohtsuki, and M. Taki
  • Journal Title: Peptide Science 2010 Pages: 113-113
  • Peer Reviewed
• [Journal Article] Synthesis of cyclic peptide/protein using the NEXT-A Reaction followed by cyclization (2011)
  • Author(s): Hamamoto, M. Sisido, T. Ohtsuki, and M. Taki
  • Journal Title: Chem. Commun Volume: 47 Pages: 9116-9116
  • Peer Reviewed
• [Journal Article] Synthesis of cyclic peptides using the NEXT-A Reaction (2011)
  • Author(s): T.Hamamoto, M.Sisido, T.Ohtsuki, M.Taki
  • Journal Title: Peptide Science 2010 Pages: 113-113
  • Peer Reviewed
• [Journal Article] Synthesis of cyclic peptide/protein using the NEXT-A Reaction followed by cyclization (2011)
  • Author(s): T.Hamamoto, M.Sisido, T.Ohtsuki, M.Taki
  • Journal Title: Chem.Commun. Volume: 47 Pages: 9116-9118
  • Peer Reviewed
• [Journal Article] Fission yeast Ubr1 ubiquitin ligase influences the oxidative stress response via degradation of active Pap1 bZIP transcription factor in the nucleus. (2011)
  • Author(s): Kitamura K, Taki M, Tanaka N, Yamashita I.
  • Journal Title: Mol Microbiol. Volume: (印刷中)
  • Peer Reviewed
• [Presentation] Exploration of Bioactive Peptides from Random Peptide Library Expressed in Yeast Cells (2013)
  • Author(s): K. Fukunaga and M. Taki
  • Organizer: 4th Asia-Pacific International Peptide Symposium (APIPS 2013)
  • Place of Presentation: Osaka
  • Year and Date: 2013-11-07
• [Presentation] 創薬システムエンジニアリング : NEXT-A 法および10BASEd-T 法の開発 (2013)
  • Author(s): 瀧 真清
  • Organizer: 群馬大学H25年度夏期複合材料研究会
  • Place of Presentation: 草津市
  • Year and Date: 2013-07-21
  • Invited
• [Presentation] NEXT-A 反応に最適な基質ペプチドの探索 (2013)
  • Author(s): 川口 淳・瀧 真清
  • Organizer: 日本化学会第93春季年会(2013)
  • Place of Presentation: 滋賀県草津市
  • Year and Date: 2013-03-22
• [Presentation] 創薬システムエンジニアリング：NEXT-A法および10BASEd-T法の開発 (2013)
  • Author(s): 瀧 真清*
  • Organizer: 群馬大学 H25年度夏期複合材料研究会
  • Place of Presentation: 草津市
  • Invited
• [Presentation] Exploration of Bioactive Peptides from Random Peptide Library Expressed in Yeast Cells (2013)
  • Author(s): Keisuke Fukunaga* and Masumi Taki
  • Organizer: 4th Asia-Pacific International Peptide Symposium (APIPS 2013)
  • Place of Presentation: Osaka
• [Presentation] NEXT-A 反応による蛋白質/ペプチドの迅速標識 (2012)
  • Author(s): 瀧 真清
  • Organizer: ハイペップ研究所第11回紅葉ワークショップ
  • Place of Presentation: 京都市
  • Year and Date: 2012-12-21
  • Invited
• [Presentation] Regiospecific modifications of (bio)macromolecules (2011)
  • Author(s): M.Taki
  • Organizer: デンマーク大学内セミナー
  • Place of Presentation: コペンハーゲン市(デンマーク)(招待講演)
  • Year and Date: 2011-09-05
• [Presentation] Synthesis of cyclic peptides/proteins using the NEXT-A cyclization reaction (2011)
  • Author(s): M.Taki, T.Hamamoto, M.Sisido
  • Organizer: 4th European Conference on Chemistry for Life Sciences
  • Place of Presentation: ブダペスト市(ハンガリー)(4ECCLS;公募型シンポ採択英語口頭発表)
  • Year and Date: 2011-09-01
• [Presentation] Introduction of functional amino acids at the N-terminus of peptide/protein by the NEXT-A (N-Terminal EXtension with Transferase and ARS) reaction (2010)
  • Author(s): M.Taki
  • Organizer: 沖縄国際ワークショップ公開講演会
  • Place of Presentation: 那覇市
  • Year and Date: 2010-07-09
• [Book] 第2章「創薬システムエンジニアリング:NEXT-A 法および10BASEd-T法の開発」, 総合コミュニケーション科学シリーズ ユニーク&エキサイティング サイエンス2 (2013)
  • Author(s): 瀧 真清
  • Publisher: 近代科学社
• [Book] 酵素利用技術大系 (2010)
  • Author(s): 瀧真清、宍戸昌彦
  • Total Pages: 8
  • Publisher: NTS社
• [Remarks]
  • URL: http://tkl.pc.uec.ac.jp/
• [Remarks] 電気通信大学 瀧研究室
  • URL: http://tkl.pc.uec.ac.jp/index.html