X-ray crystallographic studies of intr- and inter-cellular transport

2013 Fiscal Year Final Research Report

• Project/Area Number: 21227003
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: University of Hyogo
• Principal Investigator: TSUKIHARA Tomitake 兵庫県立大学, 生命理学研究科, 特任教授 (00032277)
• Co-Investigator(Kenkyū-buntansha): YAMASHITA Eiki 大阪大学, たんぱく質研究所, 助教 (00294132) ; TANAKA Hideaki 大阪大学, たんぱく質研究所, 助教 (40346169)
• Project Period (FY): 2009-05-11 – 2014-03-31
• Keywords: X線結晶解析 / 膜蛋白質 / 蛋白質 / 超分子 / 輸送

Research Abstract

Three kinds of structural research relating to the transportation were carried out. The first is the nuclear export mechanism of pre-miRNA. The second is structural study of vault that carries materials in the cell. The third is structural-functional studies of gap junction channels.
Structures of Exportin-5:RanGTP:Pre-miRNA complex, Exportin-5:RanGTP complex and Exportin-5 were determined at respective 2.9, 3.0 and 3.5 A resolutions. Comparing structures of exportin-5s in three different states and analyzing interactions between Pre-miRNA and Exportin-5, we elucidated Pre-miRNA transportation mechanism by Exportin-5:RanGTP complex. We have determined the 39-fold symmetry for the MVP-vault in addition to the vault from rat liver. Consequently, the 39-fold symmetry of vault was confirmed. We elucidated docking selectivity rules for hemichannel pairs by homology modeling based on the connexin-26 gap junction channel and functional analyses.

Research Products

• [Journal Article] New features of vault architecture and dynamic srevealed by novel refinement using the deformable elastic network approach (2013)
  • Author(s): Casanas A, Querol-Audi J, Guerra P, Pous J, Tanaka H, Tsukihara T, Verdaguer N, Fita I
  • Journal Title: Acta Crystallogr D Biol Crystallogr Volume: 69 Pages: 1054-1061
  • DOI: 10.1107/S0907444913004472
  • Peer Reviewed
• [Journal Article] A gap junction docking mechanism revealed by functional rescue of a human disease-linked connexin mutant (2013)
  • Author(s): Gong, X.Q., Nakagawa, S., Tsukihara, T. and Bai, D
  • Journal Title: J Cell Sci Volume: 126 Pages: 3113-3120
  • DOI: 10.1242/jcs.123430
  • Peer Reviewed
• [Journal Article] Structural studies of large nucleoprotein particles, vaults (2012)
  • Author(s): Tanaka H, Tsukihara T
  • Journal Title: Proc Jpn Acad Ser BPhys Biol Sci Volume: 88 Pages: 416-33
  • Peer Reviewed
• [Journal Article] Aspartic Acid Residue D3 Critically Determines Cx50 Gap Junction Channel Transjunctional Voltage-Dependent Gating and Unitary Conductance (2012)
  • Author(s): Xin L, Nakagawa S, Tsukihara T, Bai D
  • Journal Title: Biophysical Journal 2012 Volume: 102 Pages: 1022-1031
  • DOI: 10.1016/j.bpj
  • Peer Reviewed
• [Journal Article] Asparagine175 of connexin32 is a critical residue for docking and forming functional heterotypic gap junction channels with connexin26 (2011)
  • Author(s): Nakagawa S, Gong XQ, Maeda S, Dong Y, Misumi Y, Tsukihara T, Bai D
  • Journal Title: J Biol Chem Volume: 286 Pages: 19672-19681
  • DOI: 10.1074/jbc.M110.204958
  • Peer Reviewed
• [Journal Article] Structure of the gap junction channel and its implications for its biological functions (2011)
  • Author(s): Maeda S, Tsukihara T
  • Journal Title: Cell Mol Life Sci Volume: 68 Pages: 1115-29
  • DOI: 10.1007/s00018-010-0551-z
  • Peer Reviewed
• [Journal Article] Structural and functional studies of gap junction channels (2011)
  • Author(s): Nakagawa S1, Maeda S, Tsukihara T
  • Journal Title: Current Opinion in Structural Biology Volume: 20 Pages: 423-430
  • DOI: 10.1016/j.sbi.2010.05.003
  • Peer Reviewed
• [Journal Article] A high resolution structure of the pre-microRNA nuclear export machinery (2009)
  • Author(s): Okada C, Yamashita E, Lee SJ, Shibata S, Katahira J, Nakagawa A, Yoneda Y, Tsukihara T
  • Journal Title: Science Volume: 32 Pages: 1275-1279
  • DOI: 10.1126/science.1178705
  • Peer Reviewed
• [Journal Article] Selective nuclear export mechanism of small RNAs
  • Author(s): Lee SJ, Jiko C, Yamashita E, Tsukihara T
  • Journal Title: Curr Opin Struct Biol Volume: 21 Pages: 101-8
  • DOI: 10.1016/j.sbi.2010.11.004
  • Peer Reviewed
• [Presentation] 複雑さと精緻さを追求する構造生命科学/Structural life science (2013)
  • Author(s): 月原冨武
  • Organizer: 第13回日本蛋白質科学会年会
  • Place of Presentation: とりぎん文化会館(鳥取県鳥取市)
  • Year and Date: 2013-06-12
  • Invited
• [Presentation] X線結晶構造解析による生体超分子の構造形成と作動機構の研究 (2013)
  • Author(s): 月原冨武
  • Organizer: 2012年度内藤記念科学振興財団贈呈式
  • Place of Presentation: 日本工業倶楽部(東京都千代田区)
  • Year and Date: 2013-03-19
• [Presentation] Structure, functions and structural organization of gap junction channels (2013)
  • Author(s): Tomitake Tsukihara
  • Organizer: Nagoya Symposium Frontiers inStructural Physiology
  • Place of Presentation: 名古屋大学豊田講堂(愛知県名古屋市)
  • Year and Date: 2013-01-22
  • Invited
• [Presentation] Attempt to study structural analysis of disordered region using the low resolution data (2012)
  • Author(s): Kiyohito Kihira, Jiyoung Kang, Shoji Maeda, So Nakagawa, Yuko Misumi, Michihiro Suga, Eiki Yamashita, Masaru Tateno, Tomitake Tsukihara
  • Organizer: AsCA12/ CRYSTAL28 Adelaide Convention Centre
  • Place of Presentation: Adelaide, Australia
  • Year and Date: 20121202-06
• [Presentation] Structural organization of gap junction channels So Nakagawa (2012)
  • Author(s): X. Q. Gong, Shoji Maeda, Yuko Misumi, Kiyohito Kihira, Donglin Bai and Tomitake Tsukihara
  • Organizer: AsCA12/ CRYSTAL28 Adelaide Convention Centre
  • Place of Presentation: Adelaide, Australia
  • Year and Date: 20121202-06
• [Presentation] Structure and Functions of Gap Junction Channels (2012)
  • Author(s): Tomitake Tsukihara
  • Organizer: 2012 Proceedings of Fall International Convention of The Pharmaceutical Society of Korea Leadership of Pharmaceutical Scientists for Human Prosperity AW Convention Center
  • Place of Presentation: Seoul Korea
  • Year and Date: 20121023-24
  • Invited
• [Presentation] Structure and functions of the gap junction channel (2012)
  • Author(s): Tomitake Tsukihara
  • Organizer: Cold Spring Harbor Conferences Asia, Membrane Proteins : Structure and Function the Suzhou Dushu Lake Conference Center
  • Place of Presentation: Suzhou China
  • Year and Date: 20120516-20
  • Invited
• [Presentation] Structural Studies of Macromolecular Assemblies Playing Roles in Transportation Assemblies (2011)
  • Author(s): Tomitake Tsukihara
  • Organizer: XXII Congress and General Assembly of the International Union of Crystallography (IUCr2011) PALACIO MUNICIPAL DE CONGRESOS Madrid
  • Place of Presentation: Spain
  • Year and Date: 20110822-30
  • Invited
• [Presentation] Structural studies of connexin-26 gap junction channel (2011)
  • Author(s): TomitakeTsukihara
  • Organizer: 36th FEBS CONGRESS, Lingotto Conference Centre
  • Place of Presentation: TorinoItaly
  • Year and Date: 20110625-30
  • Invited
• [Presentation] ギャップ結合チャネルの構造とその働き (2011)
  • Author(s): 月原冨武
  • Organizer: 日本生化学会北陸支部第29回大会
  • Place of Presentation: 金沢大学 角間キャンパス (石川県金沢市)
  • Year and Date: 2011-05-28
  • Invited
• [Presentation] The structure of rat liver vault at 3.5 A resolution (2010)
  • Author(s): 加藤公児
  • Organizer: The 10th Conference of the Asian Crystallographic Association Bexco
  • Place of Presentation: Busan, Korea
  • Year and Date: 20101031-1103
• [Presentation] Structure and function of gap junction channels Shoji Maeda Gordon Research Conferences (2010)
  • Organizer: Ligand Recognition & Molecular GatingIl Ciocco Hotel and Resort
  • Place of Presentation: Lucca (Barga), Italy
  • Year and Date: 20100516-21
• [Presentation] The mechanisms of self-assembly of the vault : the largest cytoplasmic ribonucleoprotein complex (2010)
  • Author(s): 田中秀明
  • Organizer: Pacifichem 2010
  • Place of Presentation: Hawaii Convention Center Hawaii USA
  • Year and Date: 2010-12-17
• [Presentation] 生体超分子の構造と機能の解明 (2010)
  • Author(s): 月原冨武
  • Organizer: 日本結晶学会60周年記念
  • Place of Presentation: 大阪大学コンベンションセンターMOホール(大阪府吹田市)
  • Year and Date: 2010-12-04
• [Presentation] X-ray crystal structure of rat vault, a large nucleoprotein complex at 3.5 A resolution (2010)
  • Author(s): 田中秀明
  • Organizer: Proceedings of the International Symposium on New Drug Development Chungbuk National University
  • Place of Presentation: Chungcheongbuk-do Korea
  • Year and Date: 2010-11-18
  • Invited
• [Presentation] タンパク質合成を抑制する小さなRNAを輸送する仕組みの解明 (2010)
  • Author(s): 山下栄樹
  • Organizer: 第2回SPring-8合同コンファレンス
  • Place of Presentation: 東京ステーションコンファレンス(東京都千代田区)
  • Year and Date: 2010-11-04
  • Invited
• [Presentation] Structure and function of Connexin26 gap junction channel (2010)
  • Author(s): Tomitake Tsukihara
  • Organizer: Gordon Research Conferences, Ligand Recognition & Molecular Gating
  • Place of Presentation: Il Ciocco Hotel and Resort, Lucca, Barga Italy
  • Year and Date: 2010-05-18
• [Presentation] Structure of the Gap Junction Channel and its Implications on its Biological Functions (2010)
  • Author(s): Tomitake Tsukihara
  • Organizer: EMBO Conference Series, Catalytic mechanisms by biological systems : at the interface between chemistry and biology EMBL
  • Place of Presentation: Hamburg, Germany
  • Year and Date: 2010-05-05
• [Presentation] The structure of rat liver vault (2009)
  • Author(s): Tomitake Tsukihara
  • Organizer: AsCA '09 Joint conference of the Asian crystallographic society and Chinese crystallography society Jingyi Hotel
  • Place of Presentation: Beijing, China
  • Year and Date: 20091022-25
• [Presentation] The structure of human connexin 26 gap junction channel Shoji Maeda (2009)
  • Organizer: AsCA '09 Joint conference of the Asian crystallographic society and Chinese crystallography society Jingyi Hotel
  • Place of Presentation: Beijing, China
  • Year and Date: 20091022-25
• [Presentation] Structural insight into heterotypic channel compatibility among connexins by homology modeling So Nakagawa (2009)
  • Organizer: International gap junction conference Sedona
  • Place of Presentation: Arizona USA
  • Year and Date: 20090725-30
• [Presentation] Structure of human gap junction channel (2009)
  • Organizer: International Gap Junction Conference 2009 Shoji Maeda Sedona
  • Place of Presentation: Arizona USA
  • Year and Date: 20090725-30
• [Remarks] 月原研究室ホームページ <WEB>
  • URL: http://www.sci.u-hyogo.ac.jp/life/GCOE/japanese/pico_intro/tsukihara/index.html
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  • URL: http://youtu.be/6bxAhPQ7m24
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  • URL: http://www.protein.osaka-u.ac.jp/olabb/tsukihara/mvp/index.html
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