Structure of photoreceptor membrane protein by In-situ photoirradiated solid-state NMR

• Project/Area Number: 22770101
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: Yokohama National University
• Principal Investigator: KAWAMURA Izuru 横浜国立大学, 工学研究院, 助教 (20452047)
• Co-Investigator(Renkei-kenkyūsha): NAITO Akira 横浜国立大学, 工学研究院, 教授 (80172245)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2011: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2010: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: 固体NMR / 膜タンパク質 / 構造生命科学 / バクテリオロドプシン / 光中間体 / レチナール / センサリーロドプシン / 細胞膜 / 脂質二重膜 / 光異性化

Research Abstract

In situ photoirradiated solid-state NMR have been developed and demonstrated to successfully observe the M-photointermediate of pharaonis phoborhodopsin(ppR) and T204A mutant in the lipid environment. The C^<13> NMR signals of[20-C^<13>] retinal-ppR with transducer revealed that multiple M-photointermediates with 13-cis, 15-anti retinal configuration under continuously photoirradiated condition. Our results provide insights into the process of photocycle in the ppR with transducer protein.

Research Products

• [Journal Article] Local Dynamics Change of Bacteriorhodopsin with Retinal Isomerization under Pressure as studied by Fast Magic Angle Spinning NMR (2012)
  • Author(s): I. Kawamura, S. Yamaguchi, H. Nishikawa, K. Tajima, M. Horigome, S. Tuzi, H. Saito, A. Naito
  • Journal Title: Polym. J. Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] 動的核分極DNPによる超高感度固体NMR法の生体分子への展開 (2012)
  • Author(s): 川村出、内藤晶
  • Journal Title: 分光研究 Volume: 61巻
  • NAID: 10030318747
  • Peer Reviewed
• [Journal Article] 動的核分極DNPによる超高感度固体NMR法の生体分子への展開 (2012)
  • Author(s): 川村出
  • Journal Title: 分光研究 Volume: 61(印刷中)
  • NAID: 10030318747
  • Peer Reviewed
• [Journal Article] An Active Photoreceptor Intermediate Revealed by In Situ Photoirradiated Solid-State NMR Spectroscopy (2011)
  • Author(s): Y. Tomonaga, T. Hidaka, I. Kawamura, T. Nishio, K. Ohsawa, T. Okitsu, A. Wada, Y. Sudo, N. Kamo, A. Ramamoorthy, A. Naito
  • Journal Title: Biophys. J. Volume: 101
  • Peer Reviewed
• [Journal Article] Site-Specific Solid-State NMR Detection of Hydrogen-Deuterium Exchange Reveals Conformational Changes in a 7-Helical Transmembrane Protein (2011)
  • Author(s): S. Wang, L. Shi, I. Kawamura, L. S. Brown, V. Ladizhansky
  • Journal Title: Biophys. J. Volume: 101
  • Peer Reviewed
• [Journal Article] Conformation of a Seven Helical Transmembrane Photosensor in the Lipid Environment (2011)
  • Author(s): L. Shi, I. Kawamura, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Journal Title: Angew. Chem. Int. Volume: 50 Pages: 1302-1305
  • Peer Reviewed
• [Journal Article] Site-Specific Solid-State NMR Detection of Hydrogen-Deuterium Exchange Reveals Conformational Changes in a 7-Helical Transmembrane Protein (2011)
  • Author(s): S.Wang, L.Shi, I.Kawamura, L.S.Brown, V.Ladizhansky
  • Journal Title: Biophys.J. Volume: 101 Issue: 3 Pages: L23-L25
  • DOI: 10.1016/j.bpj.2011.06.035
  • Peer Reviewed
• [Journal Article] An active photoreceptor intermediate revealed by in situ photoirradiated solid-state NMR spectroscopy (2011)
  • Author(s): Tomonaga, Y., Hidaka, T, Kawamura, I, Nishio, T, Ohsawa, K, Okitsu, T, Wada, A, Sudo, Y, Kamo, N, Ramamoorthy, A, Naito, A.
  • Journal Title: Biophys.J. Volume: 101 Issue: 10 Pages: L50-L52
  • DOI: 10.1016/j.bpj.2011.10.022
  • Peer Reviewed
• [Journal Article] Conformation of a Seven-Helical Transmembrane Photosensor in the Lipid Environment (2011)
  • Author(s): L.Shi, I.Kawamura, K.H.Jung, L.S.Brown, V.Ladizhansky
  • Journal Title: Angew.Chem.Int.Ed. Volume: 50 Pages: 1302-1305
  • Peer Reviewed
• [Presentation] 固体NMRによる脂質二重膜中の7本膜貫通型-光受容膜タンパク質の構造解析 (2011)
  • Author(s): 川村出, L. Shi, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2011-09-22
• [Presentation] 固体NMRによる光受容膜タンパク質の構造解析 (2011)
  • Author(s): 川村出
  • Organizer: 奈良先端大・青学大・横浜国大光応答分子材料に関する3大学共同セミナー
  • Place of Presentation: 横浜国立大学中央図書館メディアホール、横浜
  • Year and Date: 2011-07-28
• [Presentation] 固体NMRによる光受容膜タンパク質の構造解析 (2011)
  • Author(s): 川村出
  • Organizer: 奈良先端大・青学大・横浜国大/光応答分子材料に関する3大学共同セミナー
  • Place of Presentation: 横浜国立大学中央図書館メディアホール、横浜
  • Year and Date: 2011-07-28
• [Presentation] Photo-induced dynamics change of Phoborhodopsin with transducer studied by C^<13> solid-state NMR (2011)
  • Author(s): I. Kawamura, R. Furusato, T. Hidaka, T. Okitsu, A. Wada, N. Kamo, A. Naito
  • Organizer: ISNMR2011
  • Place of Presentation: Yokohama Japan
• [Presentation] Dynamics change of phoborhodopsin with its cognate transducer protein as studied by in-situ photo-irradiated solid-state NMR (2011)
  • Author(s): I. Kawamura, R. Furusato, T. Kondo, T. Okitsu, A. Wada, N. Kamo, A. Naito
  • Organizer: The 4th Asia-Pacific NMR symposium
  • Place of Presentation: Beijing, China
• [Presentation] Photo-induced dynamics change of phoborhodopsin with transducer protein as studied by in-situ phoro irradiated solid-state NMR (2011)
  • Author(s): 川村出、古里龍太郎、柴藤祐介、日高徹郎、沖津貴志、和田昭盛、加茂直樹、内藤晶
  • Organizer: 第49回日本生物物理学会年会
  • Place of Presentation: 兵庫県立大学姫路書写キャンパス、姫路
• [Presentation] 固体NMRによるアミロイド.膜タンパク質の構造解析 (2011)
  • Author(s): 川村出
  • Organizer: 第12回若手NMR研究会
  • Place of Presentation: 琵琶湖リゾートクラブ、滋賀
• [Presentation] 固体NMRによる脂質膜中の7本膜貫通型-光受容膜タンパク質の構造解析 (2011)
  • Author(s): 川村出, L. Shi, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Organizer: 第11回日本蛋白質科学会年会
  • Place of Presentation: ホテル阪急エキスポパーク、大阪
• [Presentation] 固体NMRによる7本膜貫通型-光受容膜タンパク質の構造解析 (2011)
  • Author(s): 川村出, L. Shi, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Organizer: 日本化学会第91回春季年会
  • Place of Presentation: 奈川大学横浜キャンパス
• [Presentation] Photo-induced dynamics change of Phoborhodopsin with transducer as studied by ^<13>C solid-state NMR (2011)
  • Author(s): I.Kawamura, R.Furusato, T.Hidaka, T.Okitsu, A.Wada, N.Kamo, A.Naito
  • Organizer: The International Symposium on Nuclear Magnetic Resonance 2011/The 50th Memorial Annual Meeting of the Nuclear Magnetic Resonance Society of Japan
  • Place of Presentation: 大さん橋ホール、横浜
• [Presentation] Dynamics change of phoborhodopsin with its cognate transducer protein as studied by in-situ photo-irradiated solid-state NMR (Oral Presentation) Travel Award (2011)
  • Author(s): I.Kawamura, R.Furusato, T.Kondo, T.Okitsu, A.Wada, N.Kamo, A.Naito
  • Organizer: The 4th Asia-Pacific NMR symposium
  • Place of Presentation: Beijing、China(受賞)
• [Presentation] 固体NMRによる脂質二重膜中の7本膜貫通型-光受容膜タンパク質の構造解析 (2011)
  • Author(s): I.Kawamura, L.Shi, K.H.Jung, L.S.Brown, V.Ladizhansky
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館、京都
• [Presentation] Photo-induced dynamics change of Phoborhodopsin with transducer protein as studied by in-situ photo irradiated solid-state NMR (2011)
  • Author(s): 川村出、古里龍太郎、柴藤祐介、日高徹郎、沖津貴志、和田昭盛、加茂直樹、内藤晶
  • Organizer: 第49回日本生物物理学会年会
  • Place of Presentation: 兵庫県立大学姫路書写キャンパス、姫路
• [Presentation] 固体NMRによるアミロイド・膜タンパク質の構造解析 (2011)
  • Author(s): 川村出
  • Organizer: 第12回若手NMR研究会
  • Place of Presentation: 琵琶湖リゾートホテル、滋賀(招待講演)
• [Presentation] 固体NMRによる脂質膜中の7本膜貫通型-光受容膜蛋白質の構造解析 (2011)
  • Author(s): I.Kawamura, L.Shi, K.H.Jung, L.S.Brown, V.Ladizhansky
  • Organizer: 第11回日本蛋白質科学会年会
  • Place of Presentation: ホテル阪急エキスポパーク、大阪
• [Presentation] Conformation of a Seven-Helical Transmembrane Photosensor in the Lipid Environment (2011)
  • Author(s): L.Shi, I.Kawamura, S.Wang, K.H.Jung, L.S.Brown, V.Ladizhansky
  • Organizer: The 52nd ENC (Experimental Nuclear Magnetic Resonance Conference)
  • Place of Presentation: Asilomar Conference Center、California, USA
• [Presentation] 固体NMRによる7本膜貫通型-光受容膜タンパク質の構造解析 (2011)
  • Author(s): 川村出, L.Shi, L.S.Brown, V.Ladizhansky
  • Organizer: 日本化学会 第91回春季年会
  • Place of Presentation: 神奈川大学 横浜キャンパス
• [Presentation] Protein backbone conformations of archaeal-type rhodopsins as studied by solid-state NMR (2010)
  • Author(s): I. Kawamura, M. Horigome, S. Tuzi, A. Naito
  • Organizer: Pacifichem2010
  • Place of Presentation: Hawaii, Honolulu
  • Year and Date: 2010-12-16
• [Presentation] Protein-backbone conformations of archaeal-type rhodopsin as studied by solid-state NMR (2010)
  • Author(s): I.Kawamura, M.Horigome, S.Tuzi, A.Naito
  • Organizer: Pacifichem 2010
  • Place of Presentation: Hawaii Convention Center, Honolulu, Hawaii, USA(一般英語口頭発表)
  • Year and Date: 2010-12-16
• [Presentation] Three-dimensional Solid-state NMR study of Anabaena Sensory Rhodopsin in the lipid environment-Chemical Shift Assignments- (2010)
  • Author(s): I.Kawamura, L.Shi, L.S.Brown, V.Ladizhansky
  • Organizer: 第48回日本生物物理学会年会
  • Place of Presentation: 東北大学 川内キャンパス
  • Year and Date: 2010-09-21
• [Presentation] 固体NMRによるアナベナセンサリーロドプシンの構造解析 (2010)
  • Author(s): 川村出, L. Shi, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Organizer: 第49回NMR討論会
  • Place of Presentation: タワーホール船堀、東京
• [Presentation] Solid-state NMR structural study of anabaena sensory rhodopsin in the lipid environment (2010)
  • Author(s): I. Kawamura, L. Shi, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Organizer: 3rd Japanese-French Joint Seminar on Organic Photochromism
  • Place of Presentation: Nisseki Yokohama Hall, Yokohama, Japan
• [Presentation] Three-dimensional solid-state NMR study of Anabaena Sensory Rhodopsin in the lipid environment. Chemical shift assignments- (2010)
  • Author(s): 川村出, L. Shi, K. H. Jung, L. S. Brown, V. Ladizhansky
  • Organizer: 第48回日本生物物理学会年会
  • Place of Presentation: 東北大学川内キャンパス、仙台
• [Presentation] Structural change of Tyr residues in Bacteriorhodopsin corresponding to retinal configurations as studied by solid-state NMR (2010)
  • Author(s): I. Kawamura, M. Horigome, S. Tuzi, A. Naito
  • Organizer: 14th International Conference on Retinal Proteins(ICRP2010)
  • Place of Presentation: UCSC, Santa Cruz, San Francisco
• [Presentation] 光照射固体NMRを用いたppRのM中間体の観測と光異性化によるレチナールグループの構造変化の解析 (2010)
  • Author(s): 友永雄也、日高徹郎、川村出、和田昭盛、須藤雄気, 加茂直樹、内藤晶
  • Organizer: BMB2010第33回日本分子生物学会年会、第83回日本生化学会大会 合同大会
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] Local structural change and dynamics at Tyr residues in Bacteriorhodopsin corresponding to two isomers of retinal as revealed by solid-state NMR (2010)
  • Author(s): 堀籠美也子、川村出、辻暁、内藤晶
  • Organizer: BMB2010第33回日本分子生物学会年会、第83回日本生化学会大会 合同大会
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] 固体NMRによるアナベナセンサリーロドプシンの構造解析 (2010)
  • Author(s): I.Kawamura, L.Shi, K.H.Jung, L.S.Brown, V.Ladizhansky
  • Organizer: 第49回NMR討論会
  • Place of Presentation: タワーホール船堀
• [Presentation] 光照射NMRによるレチナールタンパク質の光中間体の観測 (2010)
  • Author(s): 内藤晶, 友永雄也、日高徹郎, 川村出、西尾拓道、大澤一弘、和田昭盛、須藤雄気、加茂直樹
  • Organizer: 第49回NMR討論会
  • Place of Presentation: タワーホール船堀
• [Presentation] 光照射固体NMRによる光受容膜タンパク質ppR-pHtrII複合体の負の走光性発現に関わるTyr174の局所構造変化の解析 (2010)
  • Author(s): 日高徹郎、友永雄也、川村出、和田昭盛、須藤雄気、加茂直樹、内藤晶
  • Organizer: 第49回NMR討論会
  • Place of Presentation: タワーホール船堀
• [Presentation] Solid-state NMR Structural Study of Anabaena Sensory Rhodopsin in the lipid environment (2010)
  • Author(s): I.Kawamura, L.Shi, K.H., Jung, L.S.Brown, V.Ladizhansky
  • Organizer: 3rd Japanese-French Joint Seminar on Organic Photochromism-Innovations in Photochromism-
  • Place of Presentation: 日石ホール,横浜
• [Presentation] 多次元固体NMRによるアナベナセンサリーロドプシンの構造解析 (2010)
  • Author(s): 川村出
  • Organizer: 第11回 若手NMR研究会
  • Place of Presentation: 検見川セミナーハウス、千葉
• [Presentation] Structural Change of Tyr residues in Bacteriorhodopsin corresponding to retinal configurations as studied by solid-state NMR (2010)
  • Author(s): I.Kawamura, M.Horigome, S.Tuzi, A.Naito
  • Organizer: 14th International Conference on Retinal Proteins (ICRP2010)
  • Place of Presentation: UCPC, Santa Cruz, San Francisco, USA
• [Presentation] ^<13>C固体NMRによる暗順応状態バクテリオロドプシンの構造解析 (2010)
  • Author(s): 川村出、堀籠美也子、辻暁, 内藤晶
  • Organizer: 第10回日本蛋白質科学会年会
  • Place of Presentation: 札幌コンベンションセンター
• [Book] 広がる核磁気共鳴NMRの世界 (2011)
  • Author(s): 川村出
  • Publisher: コロナ社
• [Book] 拡がるNMRの世界～40人のNMR研究者からの熱いメッセージ～"レチナール蛋白質の固体NMR構造解析"分担執筆 (2011)
  • Author(s): 川村出
  • Publisher: コロナ社
• [Remarks] 横浜国立大学研究者総覧
  • URL: http://er-web.jmk.ynu.ac.jp/
• [Remarks] 横浜国立大学研究者総覧
  • URL: http://er-web.jmk.ynu.ac.jp
• [Remarks] 横浜国立大学 研究者総覧
  • URL: http://er-web.jmk.ynu.ac.jp/