Development of a ribosomal translation system that enables efficient incorporation of nonproteinogenic amino acids

• Project/Area Number: 18H02080
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: Katoh Takayuki 東京大学, 大学院理学系研究科(理学部), 准教授 (90567760)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2021: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2020: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: ペプチド医薬 / 翻訳 / tRNA / リボソーム / 非天然アミノ酸 / 翻訳合成 / ペプチド創薬

Outline of Final Research Achievements

We developed a methodology that enables efficient ribosomal incorporation of nonproteinogenic amino acids, such as D-amino, β-amino, γ-amino, and N-methyl-amino acids, into nascent peptides. By means of this system, we constructed macrocyclic random peptide libraries bearing those nonproteinogenic amino acids and applied them to mRNA display-based in vitro selection against various disease-related target proteins. The obtained macrocyclic peptides exhibited extremely high binding affinity, inhibitory activity, and serum stability.

Academic Significance and Societal Importance of the Research Achievements

D-アミノ酸やβ-アミノ酸、γ-アミノ酸、N-メチルアミノ酸等の非天然アミノ酸はペプチドの安定性や標的への結合力、細胞膜透過性等の特性を向上させることができる。これらの非天然アミノ酸を含む特殊ペプチドの翻訳合成法を確立できれば、鋳型mRNAの配列を変えるだけで容易にランダム特殊ペプチドライブラリを構築することが可能になる。さらに、mRNAディスプレイ法などの配列スクリーニング手法を適用することで、薬理活性を持つ特殊ペプチドを効率的に探索できるため、非常に有効なペプチド創薬プラットフォームとなる。

Research Products

• [Int'l Joint Research] IECB(フランス)
• [Int'l Joint Research] IECB(フランス)
• [Journal Article] Drop-off-reinitiation triggered by EF-G-driven mistranslocation and its alleviation by EF-P (2022)
  • Author(s): Tajima Kenya、Katoh Takayuki、Suga Hiroaki
  • Journal Title: Nucleic Acids Research Volume: 50 Issue: 5 Pages: 2736-2753
  • DOI: 10.1093/nar/gkac068
  • Peer Reviewed / Open Access
• [Journal Article] In Vitro Selection of Foldamer-Like Macrocyclic Peptides Containing 2-Aminobenzoic Acid and 3-Aminothiophene-2-Carboxylic Acid (2022)
  • Author(s): Katoh Takayuki、Suga Hiroaki
  • Journal Title: Journal of the American Chemical Society Volume: 144 Issue: 5 Pages: 2069-2072
  • DOI: 10.1021/jacs.1c12133
  • Peer Reviewed
• [Journal Article] Consecutive Ribosomal Incorporation of α-Aminoxy/α-Hydrazino Acids with D/L-Configurations into Nascent Peptide Chains (2021)
  • Author(s): Katoh Takayuki、Suga Hiroaki
  • Journal Title: Journal of the American Chemical Society Volume: 143 Issue: 45 Pages: 18844-18848
  • DOI: 10.1021/jacs.1c09270
  • Peer Reviewed / Open Access
• [Journal Article] In Vitro Selection of Thioether-Closed Macrocyclic Peptide by Means of the RaPID System (2021)
  • Author(s): Katoh Takayuki、Goto Yuki、Suga Hiroaki
  • Journal Title: Methods in Molecular Biology Volume: 2371 Pages: 247-259
  • DOI: 10.1007/978-1-0716-1689-5_13
  • ISBN: 9781071616888, 9781071616895
  • Peer Reviewed
• [Journal Article] Uniform affinity-tuning of N-methyl-aminoacyl-tRNAs to EF-Tu enhances their multiple incorporation (2021)
  • Author(s): Iwane Yoshihiko、Kimura Hiroyuki、Katoh Takayuki、Suga Hiroaki
  • Journal Title: Nucleic Acids Research Volume: 49 Issue: 19 Pages: 10807-10817
  • DOI: 10.1093/nar/gkab288
  • Peer Reviewed / Open Access
• [Journal Article] In Vitro Selection of Macrocyclic <scp>d</scp>/<scp>l</scp>-Hybrid Peptides against Human EGFR (2021)
  • Author(s): Imanishi Sayaka、Katoh Takayuki、Yin Yizhen、Yamada Mituhiro、Kawai Marina、Suga Hiroaki
  • Journal Title: Journal of the American Chemical Society Volume: 143 Issue: 15 Pages: 5680-5684
  • DOI: 10.1021/jacs.1c02593
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Macrocyclic peptides exhibit antiviral effects against influenza virus HA and prevent pneumonia in animal models (2020)
  • Author(s): Saito, M Itoh Y, Yasui F, Munakata T, Yamane D, Ozawa M, Ito R, Katoh T, Ishigaki H, Nakayama M, Shichinohe S, Yamaji K, Yamamoto N, Ikejiri A, Honda T, Sanada T, Sakoda Y, Kida H, Le TH, Kawaoka Y, Ogasawara Tsukiyama-Kohara T, Suga H, & Kohara M
  • Journal Title: Nature Comunications Volume: - Issue: 1 Pages: 2654-2654
  • DOI: 10.1038/s41467-021-22964-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ribosomal synthesis and de novo discovery of bioactive foldamer peptides containing cyclic β-amino acids (2020)
  • Author(s): Katoh Takayuki、Sengoku Toru、Hirata Kunio、Ogata Kazuhiro、Suga Hiroaki
  • Journal Title: Nature Chemistry Volume: 12 Issue: 11 Pages: 1081-1088
  • DOI: 10.1038/s41557-020-0525-1
  • Peer Reviewed
• [Journal Article] Ribosomal Elongation of Aminobenzoic Acid Derivatives (2020)
  • Author(s): Katoh Takayuki、Suga Hiroaki
  • Journal Title: Journal of the American Chemical Society Volume: 142 Issue: 39 Pages: 16518-16522
  • DOI: 10.1021/jacs.0c05765
  • Peer Reviewed
• [Journal Article] Ribosomal Elongation of Cyclic γ-Amino Acids using a Reprogrammed Genetic Code (2020)
  • Author(s): Katoh Takayuki、Suga Hiroaki
  • Journal Title: Journal of the American Chemical Society Volume: 142 Issue: 11 Pages: 4965-4969
  • DOI: 10.1021/jacs.9b12280
  • Peer Reviewed
• [Journal Article] Flexizyme-catalyzed synthesis of 3'-aminoacyl-NH-tRNAs. (2019)
  • Author(s): Katoh T, Suga H.
  • Journal Title: Nucleic Acids Res. Volume: pii Issue: 9 Pages: e54-e54
  • DOI: 10.1093/nar/gkz143
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Engineering Translation Components Improve Incorporation of Exotic Amino Acids (2019)
  • Author(s): Katoh Takayuki、Suga Hiroaki
  • Journal Title: International Journal of Molecular Sciences Volume: 20 Issue: 3 Pages: 522-522
  • DOI: 10.3390/ijms20030522
  • Peer Reviewed / Open Access
• [Journal Article] Ribosomal Incorporation of Consecutive beta-Amino Acids. (2018)
  • Author(s): Katoh T, Suga H.
  • Journal Title: J Am Chem Soc. Volume: 140 Issue: 38 Pages: 12159-12167
  • DOI: 10.1021/jacs.8b07247
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] In vitro expression of genetically encoded non-standard peptides consisting of exotic amino acid building blocks. (2018)
  • Author(s): Hirose H, Tsiamantas C, Katoh T, Suga H.
  • Journal Title: Curr Opin Biotechnol. 2018 Nov 16;58: Volume: 58 Pages: 28-36
  • DOI: 10.1016/j.copbio.2018.10.012
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 細胞膜透過型Helixペプチドライブラリーの構築とその薬剤候補探索への適用 (2022)
  • Author(s): 大城 裕亮・加藤 敬行・菅裕明
  • Organizer: 日本化学会
• [Presentation] In vitro selection of antibiotic peptides that inhibit the bacterial ribosome (2022)
  • Author(s): Rei Takahashi, Takayuki Katoh, Axel Innis, Hiroaki Suga
  • Organizer: 日本化学会
• [Presentation] 環状γ-アミノ酸含有特殊ペプチドライブラリの構築と薬剤候補探索への適用 (2020)
  • Author(s): 三浦敬、加藤敬行、菅裕明
  • Organizer: 日本化学会
• [Presentation] 環状β-アミノ酸を含むヘリックスペプチドライブラリを用いた薬剤候補スクリーニング (2020)
  • Author(s): 川合茉利奈、加藤敬行、菅裕明
  • Organizer: 日本化学会
• [Presentation] Ribosomal incorporation of consecutive D- and β-amino acids (2019)
  • Author(s): Katoh Takayuki、Suga Hiroaki
  • Organizer: RNA 2019
  • Int'l Joint Research
• [Presentation] Ribosomal synthesis and de novo discovery of bioactive foldamer peptides containing β- and D-amino acids (2019)
  • Author(s): Takayuki Katoh
  • Organizer: 12th International Symposium on Aminoacyl-tRNA Synthetases
  • Int'l Joint Research / Invited
• [Presentation] Ribosomal incorporation of consecutive D- and β-amino acids (2019)
  • Author(s): Takayuki Katoh, Hiroaki Suga
  • Organizer: Ribosome 2019
  • Int'l Joint Research
• [Presentation] Logical engineering of tRNA for incorporation of D- and β-amino acids into macrocyclic peptides (2018)
  • Author(s): Takayuki Katoh
  • Organizer: 4th International Conference on Circular Proteins and Peptides (ICCPP2018)
  • Int'l Joint Research / Invited
• [Presentation] Logical engineering of D-arm and T-stem of tRNA for incorporation of non-proteinogenic amino acids (2018)
  • Author(s): Takayuki Katoh, Hiroaki Suga
  • Organizer: 27th tRNA conference
  • Int'l Joint Research