ダウンロード数: 240
このアイテムのファイル:
ファイル | 記述 | サイズ | フォーマット | |
---|---|---|---|---|
acschembio.0c00145.pdf | 1.23 MB | Adobe PDF | 見る/開く |
タイトル: | Characterization of Class IB Terpene Synthase: The First Crystal Structure Bound with a Substrate Surrogate |
著者: | Stepanova, Rafaella Inagi, Hayato Sugawara, Kei Asada, Kazuya Nishi, Tomoyuki Ueda, Daijiro Yasuno, Yoko Shinada, Tetsuro Miki, Kunio Fujihashi, Masahiro https://orcid.org/0000-0002-6882-9697 (unconfirmed) Sato, Tsutomu |
著者名の別形: | 稲木, 隼人 菅原, 啓 西, 智之 上田, 大次郎 保野, 陽子 品田, 哲郎 三木, 邦夫 藤橋, 雅宏 佐藤, 努 |
発行日: | 19-Jun-2020 |
出版者: | American Chemical Society (ACS) |
誌名: | ACS Chemical Biology |
巻: | 15 |
号: | 6 |
開始ページ: | 1517 |
終了ページ: | 1525 |
抄録: | Terpene synthases (TS) are classified into two broad types, Class I and II, based on the chemical strategy for initial carbocation formation and motif sequences of the catalytic site. We have recently identified a new class of enzymes, Class IB, showing the acceptability of long (C₂₀–C₃₅) prenyl-diphosphates as substrates and no amino acid sequence homology with known TS. Conversion of long prenyl-diphosphates such as heptaprenyl-diphosphate (C₃₅) is unusual and has never been reported for Class I and II enzymes. Therefore, the characterization of Class IB enzymes is crucial to understand the reaction mechanism of the extensive terpene synthesis. Here, we report the crystal structure bound with a substrate surrogate and biochemical analysis of a Class IB TS, using the enzyme from Bacillus alcalophilus (BalTS). The structure analysis revealed that the diphosphate part of the substrate is located around the two characteristic Asp-rich motifs, and the hydrophobic tail is accommodated in a unique hydrophobic long tunnel, where the C₃₅ prenyl-diphosphate, the longest substrate of BalTS, can be accepted. Biochemical analyses of BalTS showed that the enzymatic property, such as Mg^2⁺ dependency, is similar to those of Class I enzymes. In addition, a new cyclic terpene was identified from BalTS reaction products. Mutational analysis revealed that five of the six Asp residues in the Asp-rich motifs and two His residues are essential for the formation of the cyclic skeleton. These results provided a clue to consider the application of the unusual large terpene synthesis by Class IB enzymes. |
著作権等: | This document is the Accepted Manuscript version of a Published Work that appeared in final form in 'ACS Chemical Biology', copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acschembio.0c00145 The full-text file will be made open to the public on 31 March 2021 in accordance with publisher's 'Terms and Conditions for Self-Archiving' この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 This is not the published version. Please cite only the published version. |
URI: | http://hdl.handle.net/2433/255583 |
DOI(出版社版): | 10.1021/acschembio.0c00145 |
PubMed ID: | 32227910 |
出現コレクション: | 学術雑誌掲載論文等 |
このリポジトリに保管されているアイテムはすべて著作権により保護されています。