多剤排出輸送体の構造生物学

2015 Fiscal Year Annual Research Report

• Project/Area Number: 25291011
• Research Institution: The University of Tokyo
• Principal Investigator: 石谷 隆一郎 東京大学, 理学(系)研究科(研究院), 准教授 (90361568)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: Ｘ線結晶構造解析 / 膜タンパク質 / 多剤耐性菌 / 薬物動態

Outline of Annual Research Achievements

DMT (Drug-Metabolite Transporter) ファミリー輸送体に関しては，アミノ酸排出輸送体である細菌由来YddGに関して大量発現，結晶化に成功し，回折像測定を行った．さらに，重原子置換体の作成とSIRAS法による位相決定にも成功し，2.4Å分解能での構造決定にも成功した．現在，機能解析を行い，論文を執筆中である．真核生物由来MATEに関しては，構造解析可能なレベルの量・純度・安定性を有する真核生物由来MATE 発現系の網羅的探索を行った．現在構造解析を推進している．

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

細菌由来DMTとMATEに関しては予定通り計画が進んでおり，今後構造決定と輸送メカニズム解明を目指し，特にDMTファミリーYddGに関しては構造決定にも成功している．今後，論文投稿へと進める．真核生物由来MATEに関しては，構造解析可能なレベルの量・純度・安定性を有する真核生物由来MATE 発現系の網羅的探索を進行中であり，予定通り計画が進んでいる．

Strategy for Future Research Activity

DMTファミリー輸送体YddGに関しては，外向き開状態の構造決定に至ったが，機能解析を行い，論文を完成させ投稿を目指す．真核生物由来に関しては，今後構造解析可能な系の探索を進め構造決定を目指す．

Causes of Carryover

MATEの構造解析が完了しておらず，計画実施に必要なため．

Expenditure Plan for Carryover Budget

MATEの構造解析を推進するために使用する．

Research Products

• [Journal Article] Structure and biological function of ENPP6, a choline-specific glycerophosphodiester-phosphodiesterase (2016)
  • Author(s): Morita J; Kano K; Kato K; Takita H; Sakagami H; Yamamoto Y; Mihara E; Ueda H; Sato T; Tokuyama H; Arai H; Asou H; Takagi J; Ishitani R; Nishimasu H; Nureki O; Aoki J
  • Journal Title: Sci Rep. Volume: 6 Pages: 20995
  • DOI: 10.1038/srep20995
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structure and Engineering of Francisella novicida Cas9 (2016)
  • Author(s): Hirano H; Gootenberg JS; Horii T; Abudayyeh OO; Kimura M; Hsu PD; Nakane T; Ishitani R; Hatada I; Zhang F; Nishimasu H; Nureki O
  • Journal Title: Cell Volume: 164 Pages: 950-961
  • DOI: 10.1016/j.cell.2016.01.039
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Structural Insights into Divalent Cation Modulations of ATP-Gated P2X Receptor Channels (2016)
  • Author(s): Kasuya G, Fujiwara Y, Takemoto M, Dohmae N, Nakada-Nakura Y, Ishitani R, Hattori M, Nureki O
  • Journal Title: Cell Rep. Volume: 14 Pages: 932-944
  • DOI: 10.1016/j.celrep.2015.12.087
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Outward- and inward-facing structures of a putative bacterial transition-metal transporter with homology to ferroportin (2015)
  • Author(s): Taniguchi R, Kato HE, Font J, Deshpande CN, Wada M, Ito K, Ishitani R, Jormakka M, Nureki O
  • Journal Title: Nat. Commun. Volume: 6 Pages: 8545
  • DOI: 10.1038/ncomms9545
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Crystal Structure of Staphylococcus aureus Cas9 (2015)
  • Author(s): Nishimasu H, Cong L, Yan WX, Ran FA, Zetsche B, Li Y, Kurabayashi A, Ishitani R, Zhang F, Nureki O.
  • Journal Title: Cell Volume: 162 Pages: 1113-1126
  • DOI: doi: 10.1016/j.cell.2015.08.007
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Molecular Dynamics of Channelrhodopsin at the Early Stages of Channel Opening (2015)
  • Author(s): Takemoto M, Kato HE, Koyama M, Ito J, Kamiya M, Hayashi S, Maturana AD, Deisseroth K, Ishitani R, Nureki O
  • Journal Title: PLoS One Volume: 10 Pages: 0131094
  • DOI: doi: 10.1371/journal.pone.0131094
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Atomistic design of microbial opsin-based blue-shifted optogenetics tools (2015)
  • Author(s): Kato HE, Kamiya M, Sugo S, Ito J, Taniguchi R, Orito A, Hirata K, Inutsuka A, Yamanaka A, Maturana AD, Ishitani R, Sudo Y, Hayashi S, Nureki O
  • Journal Title: Nat. Commun. Volume: 6 Pages: 7177
  • DOI: doi: 10.1038/ncomms8177
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structural basis for dynamic mechanism of nitrate/nitrite antiport by NarK (2015)
  • Author(s): Fukuda M, Takeda H, Kato HE, Doki S, Ito K, Maturana AD, Ishitani R, Nureki O
  • Journal Title: Nat. Commun. Volume: 6 Pages: 7097
  • DOI: doi: 10.1038/ncomms8097
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Crystal Structure and Activity of the Endoribonuclease Domain of the piRNA Pathway Factor Maelstrom (2015)
  • Author(s): Matsumoto, N., Sato, K., Nishimasu, H., Namba, Y., Miyakubi, K., Dohmae, N., Ishitani, R., Siomi, H., Siomi, M. C., Nureki, O
  • Journal Title: Cell Rep. Volume: 11 Pages: 366-375
  • DOI: 10.1016/j.celrep.2015.03.030
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structural Basis for the Catalytic Mechanism of DncV, Bacterial Homolog of Cyclic GMP-AMP Synthase (2015)
  • Author(s): Kato, K., Ishii, R., Hirano, S., Ishitani, R., Nureki, O
  • Journal Title: Structure Volume: 23 Pages: 843-850
  • DOI: 10.1016/j.str.2015.01.023
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structural basis for Na transport mechanism by a light-driven Na pump (2015)
  • Author(s): Kato, H. E., Inoue, K., Abe-Yoshizumi, R., Kato, Y., Ono, H., Konno, M., Hososhima, S., Ishizuka, T., Hoque, M. R., Kunitomo, H., Ito, J., Yoshizawa, S., Yamashita, K., Takemoto, M., Nishizawa, T., Taniguchi, R., Kogure, K., Maturana, A. D., Iino, Y., Yawo, H., Ishitani, R., Kandori, H., Nureki, O
  • Journal Title: Nature Volume: 521 Pages: 48-53
  • DOI: 10.1038/nature14322
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Molecular Mechanisms of Membrane Transporters (2015)
  • Author(s): Ryuichiro Ishitani
  • Organizer: The 13th Conference of the Asian Crystallographic Association
  • Place of Presentation: Science City, Kolkata, India
  • Year and Date: 2015-12-05 – 2015-12-08
  • Int'l Joint Research / Invited
• [Presentation] Structural Basis for Dynamic Mechanism of Nitrate/Nitrite Antiport by NarK (2015)
  • Author(s): Ryuichiro Ishitani
  • Organizer: GRC Mechanisms of Membrane Transport
  • Place of Presentation: Bates College, Lewiston, ME, USA
  • Year and Date: 2015-06-28 – 2015-07-03
  • Int'l Joint Research
• [Presentation] プロトンで駆動されるトランスポーターの分子機構 (2015)
  • Author(s): 石谷隆一郎
  • Organizer: 分子研研究会「膜タンパク質内部のプロトン透過を考える」
  • Place of Presentation: 岡崎コンファレンスセンター 愛知県岡崎市明大寺町伝馬８－１
  • Year and Date: 2015-04-20 – 2015-04-21
  • Invited