Functional analysis of transporters regulating translocation of energy metabolites in rice organelle

• Project/Area Number: 24580094
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Plant nutrition/Soil science
• Research Institution: Ehime University
• Principal Investigator: NOZAWA Akira 愛媛大学, プロテオサイエンスセンター, 講師 (30432800)
• Co-Investigator(Renkei-kenkyūsha): TOZAWA Yuzuru 埼玉大学, 大学院・理工学研究科, 教授 (90363267)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 糖リン酸輸送体 / ミトコンドリアキャリア / コムギ無細胞系 / リポソーム / トランスポーター / 無細胞タンパク質合成系 / オルガネラ

Outline of Final Research Achievements

We improved a method for functional analysis of transporter proteins based on liposome-supplemented cell-free system. In the new system, detection range of transport activity against non-specific background activity was improved by up-regulated synthetic efficiency of membrane proteins in cell-free reaction. By using this method, we analyzed several transporter proteins localized in plant organelle. Substrate specificity of sugar-phosphate transporters in rice and iceplant plastids and sulfate metabolite transporter in Arabidopsis plastids.

Research Products

• [Journal Article] Modifications of wheat germ cell-free system for functional proteomics of plant membrane proteins. (2014)
  • Author(s): Akira Nozawa and Yuzuru Tozawa
  • Journal Title: Methods in Molecular Biology Volume: 1072 Pages: 259-272
  • DOI: 10.1007/978-1-62703-631-3_19
  • ISBN: 9781627036306, 9781627036313
• [Journal Article] 膜輸送体タンパク質の完全インビトロ機能解析系 (2014)
  • Author(s): 野澤彰、戸澤譲
  • Journal Title: バイオサイエンスとインダストリー Volume: 72 Pages: 190-196
  • NAID: 40020102360
• [Journal Article] Characterization of the Plastidic Phosphate Translocators in the Inducible Crassulacean Acid Metabolism Plant <i>Mesembryanthemum crystallinum</i> (2013)
  • Author(s): Kore-eda, S., Nozawa, A., Okada, Y., Takashi, K., Azad, M.A.K., Ohnishi, J., Nishiyama, Y. and Tozawa, Y.
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 77 Issue: 7 Pages: 1511-1516
  • DOI: 10.1271/bbb.130174
  • NAID: 10031190461
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Presentation] プロテオリポソーム再構成系を利用したシロイヌナズナ PAPST１ホモログAt3g51870の輸送基質の解析 (2015)
  • Author(s): 名樂 仁、井上 寛之、戸澤 譲、野澤 彰
  • Organizer: 第56回日本植物生理学会年会
  • Place of Presentation: 東京農業大学世田谷キャンパス１号館・東京都・世田谷区
  • Year and Date: 2015-03-18
• [Presentation] Cell-free system for functional analysis of membrane transporters (2014)
  • Author(s): Akira Nozawa and Yuzuru Tozawa
  • Organizer: The 12th Matsuyama International Symposium on Proteo-Sciences
  • Place of Presentation: 愛媛大学南加祈念ホール・愛媛県・松山市
  • Year and Date: 2014-09-17
  • Invited
• [Presentation] Functional analysis of membrane transporters by in vitro reconstitution system (2014)
  • Author(s): Daichi Morita, Hiroyuki Inoue, Yuki Shimonosono, Hitoshi Myoraku, Taito Takabayashi, Ryoji Fujimoto, Nobuhiro Yamashita, Yusuke Okada, Shinichi Teramoto, Hironori Takeda, Akira Nozawa and Yuzuru Tozawa
  • Organizer: The 12th Matsuyama International Symposium on Proteo-Sciences
  • Place of Presentation: 愛媛大学法文学部講義棟・愛媛県・松山市
  • Year and Date: 2014-09-17
• [Presentation] 脂質組成と再構成ミトコンドリアキャリア蛋白質の膜輸送活性との関係
  • Author(s): 野澤 彰、戸澤 譲
  • Organizer: 日本農芸化学会中四国支部第３６回講演会
  • Place of Presentation: 松江
• [Presentation] コムギ無細胞系を基盤とした膜タンパク質解析系の構築
  • Author(s): 野澤 彰
  • Organizer: 岡山大学資源植物科学研究所 拠点共同研究主催 ワークショップ 「植物ホルモンとイオン輸送体の相互理解」
  • Place of Presentation: 岡山県倉敷市
  • Invited
• [Presentation] Cell-free synthesis and characterization of plastidic transporter proteins.
  • Author(s): Akira Nozawa, Shin Kore-eda, and Yuzuru Tozawa.
  • Organizer: The 10th Matsuyama International Symposium on Cell-Free Sciences.
  • Place of Presentation: 愛媛県松山市
• [Presentation] プラスチド型リン酸輸送体ファミリータンパク質の合成と機能解析
  • Author(s): 野澤 彰、是枝 晋、戸澤 譲
  • Organizer: 第7回無細胞生命科学研究会
  • Place of Presentation: 愛媛県松山市
• [Presentation] ミトコンドリアキャリアータンパク質の無細胞解析系の構築
  • Author(s): 岡田 有右、藤本 竜治、野澤 彰、戸澤 譲
  • Organizer: 第7回無細胞生命科学研究会
  • Place of Presentation: 愛媛県松山市
• [Presentation] コムギ無細胞系を利用した膜輸送体蛋白質の機能解析系の構築
  • Author(s): 野澤 彰、戸澤 譲
  • Organizer: 第54回日本植物生理学会年会
  • Place of Presentation: 岡山県岡山市
  • Invited