Elucidation of the transport mechanism of a secondary active transporter, Mhp1.

• Project/Area Number: 21370043
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyoto University
• Principal Investigator: SHIMAMURA Tatsuro 京都大学, 医学研究科, 特定講師 (90391979)
• Co-Investigator(Kenkyū-buntansha): TAKUYA Kobayashi 京都大学, 医学研究科, 講師 (20311730) ; SHIROISHI Mitsunori 九州大学, 薬学研究院, 助教 (00380527)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥14,170,000 (Direct Cost: ¥10,900,000、Indirect Cost: ¥3,270,000); Fiscal Year 2011: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2010: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2009: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
• Keywords: X線結晶解析 / 輸送体 / ヒダントイン / トランスポーター / 交互アクセスモデル / 膜タンパク質 / X線結晶構造解析 / 交互アクセス機構 / X線結晶構造解 / Mhp1

Research Abstract

Transporters transport the substrates across the membrane by the alternating access mechanism in which the proteins have three functionally distinct conformational states(outward-facing, occluded, and inward-facing). Mhp1 is a secondary active transporter belonging to the LeuT superfamily. We succeeded to determine the Mhp1 structures in these three states of the alternating access mechanism. By the comparison of these structures, the conformational changes between these states are found to be achieved by the rigid body movement of the four helices relative to the rest of the protein. The molecular basis of the alternating access mechanism should be common among t he LeuT superfamily.

Research Products

• [Journal Article] The Na+-hydantoin membrane transport protein (2012)
  • Author(s): Jackson SM, Ivanova E, Simmons K, Patching SG, Weyand S, Shimamura T, Brueckner F, Iwata S, Sharples DJ, Baldwin SA, Sansom MPS, Beckstein O, Cameron AD, Henderson PJF
  • Journal Title: Encyclopedia of Biophysics Springer
• [Journal Article] The Na+-hydantoin membrane transport protein, Mhp1 (2012)
  • Author(s): Weyand S.Shimamura T., et al
  • Journal Title: Encyclopedia of Biophysics Volume: (掲載確定)
• [Journal Article] Production of the stable human histamine H(1) receptor in Pichia pastoris for structural determination (2011)
  • Author(s): Shiroishi M, Kobayashi T, Ogasawara S, Tsujimoto H, Ikeda-Suno C, Iwata S, Shimamura T.
  • Journal Title: Methods. Volume: 55(4) Pages: 281-286
  • Peer Reviewed
• [Journal Article] Cloning, expression and purification of the anion exchanger 1 homologue from the basidiomycete Phanerochaete chrysosporium (2011)
  • Author(s): Tokuda N, Igarashi K, Shimamura T(co-1st author), Yurugi-Kobayashi T, Shiroishi M, Ito K, Sugawara T, Asada H, Murata T, Nomura N, Iwata S, Kobayashi T.
  • Journal Title: Protein. Expr. Purif. Volume: 79(1) Issue: 1 Pages: 81-7
  • DOI: 10.1016/j.pep.2011.04.006
  • NAID: 120003255719
  • Peer Reviewed
• [Journal Article] Evaluation of the Pichia pastoris expression system for the production of GPCRs for structural analysis (2011)
  • Author(s): Asada, H., Uemura, T., Yurugi-Kobayashi, T., Shiroishi, M., Shimamura, T., Tsujimoto, H., Ito, K., Sugawara, T., Nakane, T., Nomura, N., Murata, T., Haga, T., Iwata, S., Kobayashi, T
  • Journal Title: Microbial Cell Factories Volume: 10 Issue: 1 Pages: 24-24
  • DOI: 10.1186/1475-2859-10-24
  • Peer Reviewed
• [Journal Article] Crystal Structure of Glucansucrase from the Dental Caries Pathogen Streptococcus mutans (2011)
  • Author(s): Ito K, Ito S, Shimamura T(co-1st author), Weyand S, Kawarasaki Y, Misaka T, Abe K, Kobayashi T, Cameron AD, Iwata S.
  • Journal Title: J. Mol. Biol. Volume: 408(2) Issue: 2 Pages: 177-86
  • DOI: 10.1016/j.jmb.2011.02.028
  • Peer Reviewed
• [Journal Article] The Alternating Access Mechanism of Transport as Observed in the Sodium-Hydantoin Transporter Mhp1 (2011)
  • Author(s): Weyand S, Shimamura T, Beckstein O, Sansom M, Iwata S, Henderson P, Cameron A.
  • Journal Title: J. Synchrotron Rad. Volume: 18 Issue: 1 Pages: 20-23
  • DOI: 10.1107/s0909049510032449
  • Peer Reviewed
• [Journal Article] The Nucleobase-Cation-Symport-1 family of Membrane Transport Proteins (2011)
  • Author(s): Weyand S, Ma P, Beckstein O, Baldwin J, Jackson S, Suzuki S, Shimamura T, Sansom M, Iwata S, Cameron A, Baldwin S, Henderosn P.
  • Journal Title: John Wiley and Sons, Ltd Pages: 004-009
  • DOI: 10.1002/0470028637.met268
  • Peer Reviewed
• [Journal Article] ヒダントイン輸送体に見る膜輸送機構の構造基盤 (2011)
  • Author(s): 島村達郎
  • Journal Title: 生物物理 Volume: 51 Pages: 4-9
  • NAID: 10027414674
  • Peer Reviewed
• [Journal Article] Production of the stable human histamine H(1) receptor in Pichia pastoris for structural determination (2011)
  • Author(s): Shiroishi M, Kobayashi T, Shimamura T., et al
  • Journal Title: Methods Volume: 55 Issue: 4 Pages: 281-286
  • DOI: 10.1016/j.ymeth.2011.08.015
  • Peer Reviewed
• [Journal Article] The Nucleobase-Cation-Symport-1 family of Membrane Transport Proteins (2011)
  • Author(s): Weyand S.Shimamura T., et al
  • Journal Title: Handbook of Metalloproteins Volume: 4 and 5 Pages: 848-864
• [Journal Article] The Nucleobase-Cation-Symport-1 Family of Membrane Transport Proteins (2011)
  • Author(s): Weyand S, et al.
  • Journal Title: Handbook of Metalloproteins Volume: 4&5 Pages: 848-864
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of a glucansucrase from the dental caries pathogen Streptococcus mutans (2010)
  • Author(s): Ito K, Ito S, Shimamura T, Kawarasaki Y, Abe K, Misaka T, Kobayashi T, Iwata S.
  • Journal Title: Acta Crystallogr Volume: 66 Issue: 9 Pages: 1086-1088
  • DOI: 10.1107/s1744309110029714
  • Peer Reviewed
• [Journal Article] 立体構造より明らかとなったヒダントイン輸送体Mhp 1の輸送機構 (2010)
  • Author(s): 島村達郎
  • Journal Title: 実験医学28 Volume: 28 Pages: 2639-2642
• [Journal Article] Molecular Basis of Alternating Access Membrane Transport by the Sodium-Hydantoin Transporter Mhp1 (2010)
  • Author(s): Shimamura T, Weyand S, Beckstein O, Rutherford NG, Hadden JM, Sharples D, Sansom MSP, Iwata S, Henderson PJ, Cameron AD.
  • Journal Title: Science(Article) Volume: 328 Issue: 5977 Pages: 470-473
  • DOI: 10.1126/science.1186303
  • NAID: 10027414674
  • Peer Reviewed
• [Journal Article] Molecular Basis of Alternating Access Membrane Transport by the Sodium-Hydantoin Transporter, Mhp1 (2010)
  • Author(s): Shimamura T, et al.
  • Journal Title: Science Volume: 328 Pages: 470-473
  • NAID: 10027414674
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of a glucansucrase from the dental caries pathogen Streptococcus mutans (2010)
  • Author(s): Ito K, et al.
  • Journal Title: Acta Crystallographica Section F Volume: 66 Pages: 1086-1088
  • Peer Reviewed
• [Journal Article] The alternating access mechanism of transport as observed in the sodium-hydantoin transporter Mhp1. (2010)
  • Author(s): Weyand S, et al.
  • Journal Title: Journal of Synchrotron Radiation Volume: 18 Pages: 20-23
  • Peer Reviewed
• [Journal Article] Molecular Basis of Alternating Access Membrane Transport by the Sodium-Hydantoin Transporter Mhp1 (2010)
  • Author(s): 島村達郎, CameronAD, 岩田想
  • Journal Title: 実験医学 Volume: 28 Pages: 2639-2642
  • NAID: 10027414674
• [Journal Article] Molecular Basis of Alternating Access Membrane Transport by the Sodium-Hydantoin Transporter, Mhp1 (2010)
  • Author(s): Shimamura T, et al.
  • Journal Title: Science (印刷中)
  • NAID: 10027414674
  • Peer Reviewed
• [Journal Article] Crystal Structure of Glucansucrase from the Dental Caries Pathogen Streptococcus mutans.
  • Author(s): Ito K, et al.
  • Journal Title: Journal of Molecular Biology Volume: (印刷中)
  • Peer Reviewed
• [Presentation] 膜蛋白質の結晶化 (2012)
  • Author(s): 島村達郎
  • Organizer: 第12回日本蛋白質科学会
  • Place of Presentation: 名古屋国際会議場(愛知県)
  • Year and Date: 2012-06-20
• [Presentation] 膜蛋白質の結晶化 (2012)
  • Author(s): 島村達郎
  • Organizer: 第12回日本蛋白質科学会
  • Place of Presentation: 名古屋国際会議場(愛知県)(招待講演)(発表確定)
  • Year and Date: 2012-06-20
• [Presentation] Structural studies of human G-protein coupled receptors (2012)
  • Author(s): Shimamura T.
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大学(京都府)
  • Year and Date: 2012-03-25
• [Presentation] Structural studies of human G-protein coupled receptors (2012)
  • Author(s): Shimamura T.
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大学(京都府)(招待講演)
  • Year and Date: 2012-03-25
• [Presentation] 微結晶を用いた創薬ターゲット膜タンパク質の構造解析とSACLAの可能性 (2012)
  • Author(s): 島村達郎
  • Organizer: 第25回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 鳥栖市民文化会館(佐賀県)
  • Year and Date: 2012-01-09
• [Presentation] 微結晶を用いた創薬ターゲット膜タンパク質の構造解析とSACLAの可能性 (2012)
  • Author(s): 島村達郎
  • Organizer: 第25回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 鳥栖市民文化会館(佐賀県)(招待講演)
  • Year and Date: 2012-01-09
• [Presentation] 二次性能動輸送体Mhp1の交互アクセス機構の解明 (2011)
  • Author(s): 島村達郎
  • Organizer: 2011年度日本生体エネルギー研究会
  • Place of Presentation: 京都産業大学(京都府)
  • Year and Date: 2011-12-21
• [Presentation] Structural basis of secondary active transport in the sodium-hydantoin transporter Mhp1 (2011)
  • Author(s): Shimamura T.
  • Organizer: The JSPS/iCeMS International Symposium.
  • Place of Presentation: 京都大学(京都府)
  • Year and Date: 2011-11-17
• [Presentation] Structural basis of secondary active transport in the sodium-hydantoin transporter Mhp1 (2011)
  • Author(s): Shimamura T.
  • Organizer: The JSPS/iCeMS International Symposium
  • Place of Presentation: 京都大学(京都府)(招待講演)
  • Year and Date: 2011-11-17
• [Presentation] ヒト由来ヒスタミン受容体H1の構造解析戦略 (2011)
  • Author(s): 島村達郎
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都国際会館(京都府)
  • Year and Date: 2011-09-24
• [Presentation] ヒト由来ヒスタミン受容体H1の構造解析戦略 (2011)
  • Author(s): 島村達郎, ら
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都国際会館(京都府)(招待講演)
  • Year and Date: 2011-09-24
• [Presentation] ヒダントイン輸送体Mhp 1の基質輸送中の動き (2011)
  • Author(s): 島村達郎
  • Organizer: 生理研研究会
  • Place of Presentation: 岡崎
  • Year and Date: 2011-09-08
• [Presentation] ヒダントイン輸送体Mhp1の基質輸送中の動き (2011)
  • Author(s): 島村達郎
  • Organizer: 生理研研究会
  • Place of Presentation: 岡崎コンファレンスセンター(愛知県)(招待講演)
  • Year and Date: 2011-09-08
• [Presentation] Structure of the human histamine H1 receptor with doxepin (2011)
  • Author(s): Shimamura T., et al.
  • Organizer: International Union of Crystallography, XXII Congress and General Assembly
  • Place of Presentation: Madrid(Spain)
  • Year and Date: 2011-08-29
• [Presentation] Structure of the human histamine H1 receptor with doxepin (2011)
  • Author(s): Shimamura T., et al
  • Organizer: International Union of Crystallography, XXII Congress and General Assembly
  • Place of Presentation: マドリッド(スペイン)(招待講演)
  • Year and Date: 2011-08-29
• [Presentation] Membrane Proteins : the Last Frontier in Structural Biology (2010)
  • Author(s): Iwata S.
  • Organizer: Aminof Prize Lecuture at Stockholm University
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2010-03-31
• [Presentation] Structure and Mechanism of Mhp1 (2010)
  • Author(s): Cameron A.D.
  • Organizer: 第3回日英構造プロテオミクスシンポジウム
  • Place of Presentation: 横浜
  • Year and Date: 2010-02-12
• [Presentation] THREE CRYSTAL STRUCTURES REVEAL THE ALTERNATING ACCESS MECHANI M OF A SECONDARY ACTIVE TRANSPORTER (2010)
  • Author(s): Shimamura T.
  • Organizer: The 3rd Annual NIH Roadmap Meeting on Membrane Protein Technologies
  • Place of Presentation: San Diego(USA)
• [Presentation] Crystal structure of the secondary-active transporter (2010)
  • Author(s): 島村達郎
  • 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(北海道)
• [Presentation] Crystal structure of the secondary-active transporter, Mhp1. (2010)
  • Author(s): 島村達郎
  • Organizer: 第10回日本蛋白質科学会年会
  • Place of Presentation: 札幌
• [Presentation] THREE CRYSTAL STRUCTURES REVEAL THE ALTERNATING ACCESS MECHANISM OF A SECONDARY ACTIVE TRANSPORTER, MHP1 (2010)
  • Author(s): Shimamura T.
  • Organizer: The 3^<rd> Annual NIH Roadmap Meeting on Membrane Protein Technologies
  • Place of Presentation: アメリカ
• [Presentation] Crystal structure of the secondary-active transporter, Mhp1 (2010)
  • Author(s): Shimamura T, et al.
  • Organizer: 第10回日本蛋白質科学会年会
  • Place of Presentation: 札幌
• [Presentation] Structure and Function of Membrane Transporters (2009)
  • Author(s): Iwata S.
  • Organizer: Taiwan-Japan Symposium on Crystallography
  • Place of Presentation: 大阪
  • Year and Date: 2009-12-09
• [Presentation] Structure and Function of Membrane Transporters (2009)
  • Author(s): Iwata S.
  • Organizer: Seminar at the ILL Chadwick amphitheatre
  • Place of Presentation: Grenoble, France
  • Year and Date: 2009-10-28
• [Presentation] 構造研究から見えてきたトランスポーターの膜輸送機構 (2009)
  • Author(s): Iwata S.
  • Organizer: 第56回日本生化学会近畿支部特別講演
  • Place of Presentation: 大阪
  • Year and Date: 2009-07-18
• [Presentation] Towards the Molecular Mechanism of Solute Carrier Proteins (2009)
  • Author(s): Iwata S.
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
• [Presentation] Structure and Function of Membrane Transporters (2009)
  • Author(s): Iwata S.
  • Organizer: 24th JSSX Annual Meeting
  • Place of Presentation: 京都
• [Remarks]
  • URL: http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2010/100423_1.htm
• [Remarks]
  • URL: http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2010/100423_1.htm
• [Remarks]
  • URL: http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2010/100423_1.htm