Study on post-translational isoprenylation of a tryptophan residue in quorum sensing pheromone

• Project/Area Number: 24688011
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Bioproduction chemistry/Bioorganic chemistry
• Research Institution: The University of Tokyo (2014-2016); Chubu University (2012-2013)
• Principal Investigator: Okada Masahiro 東京大学, 薬学研究科(研究院), 准教授 (40377792)
• Project Period (FY): 2012-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥27,170,000 (Direct Cost: ¥20,900,000、Indirect Cost: ¥6,270,000); Fiscal Year 2015: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2013: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2012: ¥14,300,000 (Direct Cost: ¥11,000,000、Indirect Cost: ¥3,300,000)
• Keywords: クオラムセンシング / 翻訳後修飾 / 枯草菌 / トリプトファン / イソプレニル化 / フェロモン

Outline of Final Research Achievements

Post-translational isoprenylation of a tryptophan residue was first found in a prokaryotes Bacillus subtilis. In order to evaluate the diversity of post-translational isoprenylation, we focused on the modification enzymes. Based on the information of the essential amino acid residues for the isoprenylation activity, a novel Trp-farnesyltransferase and a novel Trp-farnesylated peptide were found in another phyla of bacteria. Then, X-ray crystal analysis of the Trp-farnesyltransferase was carried out, and then genome mining for proteins sharing the structural features of the enzyme revealed that post-translational isoprenylation of tryptophan was widely distributed in several phyla of bacteria.
In addition, a novel Trp-farnesylated peptide involved in the formation of sticky biofilm was found from B. subtilis subsp. natto. Then, detailed farnesylation analyses of the enzyme revealed that post-translationally isoprenylating enzyme for Trp residue exhibited broad substrate tolerance.

Research Products

• [Journal Article] Posttranslational isoprenylation of tryptophan in bacteria (2017)
  • Author(s): Masahiro Okada, Tomotoshi Sugita, Ikuro Abe
  • Journal Title: Beilstein J. Org. Chem. Volume: 13 Pages: 338-346
  • DOI: 10.3762/bjoc.13.37
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Characterization of giant modular PKSs provides insight into genetic mechanism for structural diversification of aminopolyol polyketides (2017)
  • Author(s): Zhang, L., Hashimoto, T., Qin, B., Hashimoto, J., Kozone, I., Kawahara, T., Okada, M., Awakawa, T., Ito, T., Asakawa, Y., Ueki, M., Takahashi, S., Osada, H., Wakimoto, T., Ikeda, H., Shin-ya, K., Abe, I.
  • Journal Title: Angew. Chem. Int. Ed. Volume: 56 Issue: 7 Pages: 1740-1745
  • DOI: 10.1002/anie.201611371
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Stereospecific prenylation of tryptophan by a cyanobacterial post-translational modification enzyme (2016)
  • Author(s): Okada, M., Sugita, T., Akita, K., Nakashima, Y., Tian, T., Li, C., Mori, T., Abe, I.
  • Journal Title: Org. Biomol. Chem. Volume: 14 Issue: 40 Pages: 9639-9644
  • DOI: 10.1039/c6ob01759b
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Genome-based discovery of an unprecedented cyclization mode in fungal sesterterpenoids biosynthesis (2016)
  • Author(s): Okada, M., Matsuda, Y., Mitsuhashi, T., Hoshino, S., Mori, T., Nakagawa, K., Quan, Z., Qin, B., Zhang, H., Hayashi, F., Kawaide, H., Abe, I.
  • Journal Title: J. Am. Chem. Soc. Volume: 138 Issue: 31 Pages: 10011-10018
  • DOI: 10.1021/jacs.6b05799
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Astellifadiene, a unique tetracyclic fungal sesterterpene: structure determination by an NMR-coupled crystalline sponge method and elucidation of its biosynthesis (2016)
  • Author(s): Matsuda, Y., Mitsuhashi,T., Lee, S., Hoshino, M., Mori, T., Okada, M., Zhang, H., Hayashi, F., Fujita, M., Abe, I.
  • Journal Title: Angew. Chem. Int. Ed. Volume: 55 Issue: 19 Pages: 5785-5788
  • DOI: 10.1002/anie.201601448
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Effective production of aromatic polyketides in Streptomyces bacteria using combined-culture (2016)
  • Author(s): Shotaro Hoshino, Masahiro Okada, Hiroyasu Onaka, Ikuro Abe.
  • Journal Title: Nat. Prod. Commun. Volume: 11 Pages: 979-981
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Manipulation of prenylation reactions by structure-based engineering of bacterial indolactam prenyltransferases (2016)
  • Author(s): T. Mori, L. Zhang, T. Awakawa, S. Hoshino, M. Okada, H. Morita, I. Abe
  • Journal Title: Nature Communications Volume: 7 Issue: 1 Pages: 10849-10851
  • DOI: 10.1038/ncomms10849
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] An unusual chimeric diterpene synthase from Emericella varicolor and its functional conversion into a sesterterpene synthase by domain swapping (2016)
  • Author(s): B. Qin, Y. Matsuda, T. Mori, M. Okada, Z. Quan, T. Mitsuhashi, T. Wakimoto, I. Abe
  • Journal Title: Angew. Chem. Int. Ed. Volume: 55 Issue: 5 Pages: 1658-1661
  • DOI: 10.1002/anie.201509263
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Chemical Structure and Biological Activity of a Quorum Sensing Pheromone from Bacillus subtilis subsp. natto. (2015)
  • Author(s): M. Okada, Y. Nakamura, S. Hayashi, K. Ozaki, and S. Usami.
  • Journal Title: Bioorg. Med. Chem. Lett. Volume: 25 Issue: 19 Pages: 4293-4296
  • DOI: 10.1016/j.bmcl.2015.07.083
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Dietziamides, novel tetramic acid dimers from Dietzia timorensis MZ-3 with antioxidative activity (2015)
  • Author(s): S. Hoshino, T. Wakimoto, H. Zhang, F. Hayashi, M. Okada, I. Abe
  • Journal Title: Bioorg. Med. Chem. Lett. Volume: 25 Issue: 18 Pages: 3953-3955
  • DOI: 10.1016/j.bmcl.2015.07.035
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Niizalactams A-C, multicyclic macrolactams isolated from combined culture of Streptomyces with mycolic acid-containing bacterium (2015)
  • Author(s): S. Hoshino, M. Okada, T. Wakimoto, H. Zhang, F. Hayashi, H. Onaka, I. Abe
  • Journal Title: J. Nat. Prod. Volume: 78 Issue: 12 Pages: 3011-3017
  • DOI: 10.1021/acs.jnatprod.5b00804
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Identification of a quorum sensing pheromone posttranslationally farnesylated at the internal tryptophan residue from Bacillus subtilis subsp. natto (2015)
  • Author(s): Shunsuke Hayashi, Syohei Usami, Yuta Nakamura, Koki Ozaki, Masahiro Okada
  • Journal Title: Biosci. Biotechnol. Biochem. Volume: 79 Issue: 10 Pages: 1567-1569
  • DOI: 10.1080/09168451.2015.1032884
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A region corresponding to second aspartate-rich motif in tryptophan isoprenylating enzyme, ComQ, serves as a substrate-binding site. (2014)
  • Author(s): M. Okada, A. Ishihara, R. Yamasaki, F. Tsuji, S. Hayashi, S. Usami, and Y. Sakagami.
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 印刷中 Issue: 4 Pages: 550-555
  • DOI: 10.1080/09168451.2014.891932
  • Peer Reviewed
• [Journal Article] Lack of the Consensus Sequence Necessary for Tryptophan Prenylation in the ComX Pheromone Precursor (2012)
  • Author(s): F. Tsuji, A. Ishihara, A. Nakagawa, M. Okada, S. Kitamura, K. Kanamaru, Y. Masuda, K. Murakami, K. Irie, and Y. Sakagami.
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 76 Issue: 8 Pages: 1492-1496
  • DOI: 10.1271/bbb.120206
  • NAID: 10031065914
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Creation of pure nanodrugs and their anticancer properties (2012)
  • Author(s): H. Kasai, T. Murakami, Y. Ikuta, Y. Koseki, K. Baba, H. Oikawa, H. Nakanishi, M. Okada, M. Shoji, M. Ueda, H. Imahori, M. Hashida
  • Journal Title: Angew. Chem. Int. Ed Volume: 51 Issue: 41 Pages: 10315-10318
  • DOI: 10.1002/anie.201204596
  • Peer Reviewed
• [Presentation] Post-translational prenylation of tryptophan. (2017)
  • Author(s): Masahiro Okada
  • Organizer: 日本化学会第97春季年会 International Symposium on Molecular Science
  • Place of Presentation: 慶應義塾大学 (神奈川県, 横浜市)
  • Year and Date: 2017-03-18
  • Int'l Joint Research / Invited
• [Presentation] 翻訳後修飾によるトリプトファンのイソプレニル化 (2017)
  • Author(s): 岡田 正弘
  • Organizer: 高分子・ハイブリッド材料研究センター (PHyM) 若手フォーラム
  • Place of Presentation: 東北大学 (宮城県, 仙台市)
  • Year and Date: 2017-02-27
  • Invited
• [Presentation] 翻訳後修飾によるトリプトファンのイソプレニル化 (2016)
  • Author(s): 岡田 正弘
  • Organizer: 第78回講演会 有機化学研究会（白鷺セミナー）
  • Place of Presentation: 大阪府立大学 (大阪府, 堺市)
  • Year and Date: 2016-12-06
  • Invited
• [Presentation] Post-translational prenylation of a tryptophan residue in quorum sensing pheromones. (2016)
  • Author(s): Masahiro Okada
  • Organizer: 1st International Conference on Advance Pharmacy and Pharmaceutical Sciences (ICAPPS)
  • Place of Presentation: Universitas Indonesia, Depok, Jakarta, Indonesia.
  • Year and Date: 2016-10-21
  • Int'l Joint Research / Invited
• [Presentation] 翻訳後修飾によるトリプトファンのイソプレニル化 (2016)
  • Author(s): 岡田 正弘
  • Organizer: 日本農芸化学会関東支部2016年度例会
  • Place of Presentation: 東京農業大学 (東京都, 世田谷区)
  • Year and Date: 2016-06-25
  • Invited
• [Presentation] Post-Translational Isoprenylation of Tryptophan (2015)
  • Author(s): M. Okada
  • Organizer: Pacifichem2015
  • Place of Presentation: Honolulu, USA
  • Year and Date: 2015-12-20
  • Int'l Joint Research / Invited
• [Presentation] Post-Translational Isoprenylation of Tryptophan (2015)
  • Author(s): M. Okada
  • Organizer: The Scripps Institution of Oceanography symposium
  • Place of Presentation: San Diego, USA
  • Year and Date: 2015-07-31
  • Invited
• [Presentation] 納豆菌由来のComXフェロモンの解明 (2015)
  • Author(s): 岡田正弘
  • Organizer: 生合成マシナリー第7回若手シンポジウム
  • Place of Presentation: 東京、東京工業大学
  • Year and Date: 2015-06-20
  • Invited
• [Presentation] トリプトファンイソプレニルトランスフェラーゼのメカニズム解明とマシナリーの構築. 生合成マシナリー第6回若手シンポジウム (2014)
  • Author(s): 岡田正弘
  • Organizer: 生合成マシナリー第6回若手シンポジウム
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2014-08-06
  • Invited
• [Presentation] トリプトファンイソプレニルトランスフェラーゼのメカニズム解明とマシナリーの構築. 生合成マシナリー第6回若手シンポジウム (2014)
  • Author(s): 岡田正弘
  • Organizer: 生合成マシナリー第7回公開シンポジウム
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2014-06-21
  • Invited
• [Presentation] Peptide isoprenylation (2012)
  • Author(s): M. Okada.
  • Organizer: International Symposiumon Natural Products Chemistry and Chemical Biology 2012
  • Place of Presentation: Hangzhou, China
• [Presentation] 翻訳後修飾によりトリプトファンがイソプレニル化された新規ペプチドの探索
  • Author(s): 宇佐美 翔平、林 峻輔、中村 優太、岡田 正弘
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 神奈川, 明治大学