Crystal structure of human Band3 (anion exchanger)

• Project/Area Number: 23370049
• 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: KOBAYASHI TAKUY 京都大学, 医学(系)研究科(研究院), 准教授 (20311730)
• Co-Investigator(Kenkyū-buntansha): HAMASAKI Naotaka 長崎国際大学, 薬学部, 名誉教授 (00091265)
• Project Period (FY): 2011-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000); Fiscal Year 2013: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2012: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2011: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: X線結晶構造解析 / アニオントランスポーター / バンド3 / 赤血球 / 抗体 / 結晶構造解析

Research Abstract

Human AE1 is a 110kDa glycoprotein. It is built from two domains, a cytosolic N-terminal domain (residues 1-360) and an integral membrane domain (residues 361-911). Anion exchange is catalysed by the C-terminal domain. However, only the crystal structure of the cytosolic N-terminal domain, important as an anchoring point for other proteins including the scaffolding protein ankyrin and deoxy-hemoglobin, has been determined. As such the details of the topology, the substrate recognition, and the anion-transport mechanism of this fundamental protein remain unclear. We analyzed the crystal structure at 3.4 angstrom resolution of the membrane domain of human AE1 locked in an outward open conformation by a covalently bound H2DIDS-inhibitor in complex with a Fab fragment from a monoclonal antibody. We also need the structure of an inward-facing open conformation or an outward-facing open conformation at higher resolution to understand the fully anion exchange mechanism of AE1.

Research Products

• [Journal Article] GPCRの立体構造から解き明かす生命科学 (2014)
  • Author(s): 小林拓也、岩田想
  • Journal Title: 実験医学増刊 羊土社 Volume: Vol. 32, No. 10 Pages: 84-91
• [Journal Article] Synthesis of photoreactive 2-phenethylamine derivatives Synthesis of adenosine derivative enabling functional analysis of adenosine receptors by photoaffinity labeling (2013)
  • Author(s): Murai, Y., Masuda, K., Ogasawara, Y., Wang, L., Hashidoko, Y., Hatanaka, Y., Iwata, S., Kobayashi, T., and Hashimoto, M
  • Journal Title: Eur. J. Org. Chem Volume: 2013 Pages: 2428-2433
  • NAID: 120005981508
• [Journal Article] Synthesis of photoreactive 2-phenethylamine derivatives - Synthesis of adenosine derivative enabling functional analysis of adenosine receptors by photoaffinity labeling. (2013)
  • Author(s): Murai Y. et al.
  • Journal Title: Eur. J. Org. Chem. Volume: 2013 Pages: 2428-2433
  • Peer Reviewed
• [Journal Article] G蛋白質共役受容体のX線結晶構造解析の進歩 (2013)
  • Author(s): 小林拓也
  • Journal Title: 血栓と循環 メディカルレビュー社 Volume: Vol. 21, No. 3 Pages: 33-40
  • Peer Reviewed
• [Journal Article] Structure of the human M2 muscarinic acetylcholine receptor bound to an antagonist (2012)
  • Author(s): Haga, K., Kruse, A.C., Asada, H., Yurugi-Kobayashi, T., Shiroishi, M., Zhang, C., Weis, W.I., Okada, T., Kobilka, B.K., Haga, T. and Kobayashi, T
  • Journal Title: Structure of the human M2 muscarinic acetylcholine receptor bound to an antagonist Volume: 482 Pages: 547-551
  • NAID: 120003752107
• [Journal Article] Platform for the rapid construction and evaluation of GPCRs for crystallography in Saccharomyces cerevisiae (2012)
  • Author(s): Shiroishi, M., Tsujimoto, H., Makyio, H., Asada, H., Yurugi-Kobayashi, T., Shimamura, T., Nomura, N., Haga, T., Iwata, S. and *Kobayashi, T
  • Journal Title: Microb. Cell Fact Volume: 11 Pages: 78-78
• [Journal Article] Production of the stable human histamine H1 receptor in Pichia pastoris for structural determination (2012)
  • Author(s): Shiroishi, M., Kobayashi, T., Ogasawara, S., Tsujimoto, H., Ikeda-Suno, C., Iwata, S. and Shimamura, T
  • Journal Title: Methods Volume: 55 Pages: 281-286
• [Journal Article] protein-coupled receptor inactivation by an allosteric inverse-agonist antibody (2012)
  • Author(s): Hino, T., Arakawa, T., Iwanari, H., Yurugi-Kobayashi, T., Ikeda-Suno, C., Nakada-Nakura, Y., Kusano-Arai, O., Weyand, S., Shimamura, T., Nomura, N., Cameron, A.D., Kobayashi, T., Hamakubo, T., Iwata, S. and Murata, T. G
  • Journal Title: Nature Volume: 482 Pages: 237-240
• [Journal Article] Platform for the rapid construction and evaluation of GPCRs for crystallography in Saccharoroyces cerevisiae. (2012)
  • Author(s): Shiroishi M., et al.
  • Journal Title: Micorb, Cell Fact. Volume: 11 Issue: 1 Pages: 78-78
  • DOI: 10.1186/1475-2859-11-78
  • Peer Reviewed
• [Journal Article] Structure of the human M2 muscarinic acetylcholine receptor bound to an antagonist (2012)
  • Author(s): Haga K., et al
  • Journal Title: Nature Volume: 482 Issue: 7386 Pages: 547-551
  • DOI: 10.1038/nature10753
  • Peer Reviewed
• [Journal Article] G protein-coupled receptor inactivation by an allosteric inverse-agonist antibody (2012)
  • Author(s): Hino T., et al
  • Journal Title: Nature Volume: 482 Issue: 7384 Pages: 237-40
  • DOI: 10.1038/nature10750
  • Peer Reviewed
• [Journal Article] Structure of the human histamine H1 receptor complex with doxepin (2011)
  • Author(s): Shimamura, T., Shiroishi, M., Weyand, S., Tsujimoto, H., Winter, G., Katritch, V., Abagyan, R., Cherezov, V., Liu, W., Han, G.W., Kobayashi, T., Stevens, R.C. and Iwata, S
  • Journal Title: Nature Volume: 475 Pages: 65-70
  • NAID: 120004225543
• [Journal Article] Cloning, expression and purification of the anion exchanger 1 homologue from basidiomycete Phanerochaete chrysosporium (2011)
  • Author(s): Tokuda, N., Igarashi, K., Shimamura, T., Yurugi-Kobayashi, T., Shiroishi, M., Ito, K., Sugawara, T., Asada, H., Murata, T., Nomura, N., Iwata, S. and Kobayashi, T
  • Journal Title: Protein Expr. Purif Volume: 79 Pages: 81-87
  • NAID: 120003255719
• [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. and Kobayashi, T
  • Journal Title: Microb. Cell Fact Volume: 10 Pages: 24-24
• [Journal Article] Cysteine-to-serine shuffling using a yeast expression system improves protein secretion : case of a nonglycosylated mutant of miracullin, a taste-modifying protein (2011)
  • Author(s): Ito, K., Sugawara, T., Koizumi, A., Nakajima, K., Shimizu-Ibuka, A., Shiroishi, M., Asada, H., Yurugi-Kobayashi, T., Shimamura, T., Asada, T., Misaka, T., Iwata, S., Kobayashi, T. and Abe, K
  • Journal Title: Biotech. Lett Volume: 33 Pages: 103-107
• [Journal Article] Crystal structure of Glucansucrase from the dental caries pathogen Streptococcus mutans (2011)
  • Author(s): Ito, K., Ito, S., Shimamura, T., Weyand, S., Kawarasaki, Y., Misaka, T., Abe, K., Kobayashi, T., Cameron, A.D., and Iwata S
  • Journal Title: J. Molecular Biology Volume: 408 Pages: 177-186
• [Presentation] Towards structure based GPCR pharmacology (2012)
  • Author(s): 小林拓也
  • Organizer: Gordon Research Conference
  • Place of Presentation: 米国
• [Presentation] Crystallization of human membrane proteins using antibody fragments
  • Author(s): Kobayashi, T
  • Organizer: Protein Island Matsuyama (PIM) International Symposium 2013
  • Place of Presentation: Matsuyama
  • Invited
• [Presentation] Crystallization of mammalian membrane proteins using antibody fragments
  • Author(s): Kobayashi, T
  • Organizer: 4th International Symposium on Diffraction Structural Biology
  • Place of Presentation: Nagoya
  • Invited
• [Presentation] Towards Structure Based GPCRs Pharmacology; From Knockout Mice to Crystal Structures
  • Author(s): Kobayashi, T.
  • Organizer: Gordon Research Conference
  • Place of Presentation: Venture, CA, USA
  • Invited
• [Presentation] 機能性抗体を利用したGPCRのX線結晶構造解析への試み(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 千里ライフサイエンスセミナー
  • Place of Presentation: 大阪
• [Presentation] 「GPCRの構造解析」から創薬は可能か?
  • Author(s): 小林拓也
  • Organizer: 持田製薬株式会社
  • Place of Presentation: 静岡
• [Presentation] GPCRのシグナル伝達分子機構の解明から創薬に向けて(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜
• [Presentation] GPCRのX線結晶構造解析と創薬に必要なGPCRの構造情報について(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: よこはまNMR構造生物学研究会第47回ワークショップ
  • Place of Presentation: 横浜
• [Presentation] GPCRをターゲットにしたX線結晶構造解析の現状と今後の展望(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 2012年日本神経化学会公開シンポジウム
  • Place of Presentation: 神戸
• [Presentation] GPCRをターゲットにした「構造に指南された創薬」を目指すための三つの戦略
  • Author(s): 小林拓也
  • Organizer: 構造活性フォーラム2012
  • Place of Presentation: 京都
• [Presentation] G蛋白共役受容体(GPCR)のX線結晶構造解析を成功させるための戦略(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 第85回日本薬理学会年会
  • Place of Presentation: 京都
• [Presentation] ムスカリン性アセチルコリン受容体M2サブタイプの三次元結晶構造解析に向けた試み(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
• [Presentation] Molecular Basis of Antihistamine Specificity Against Human Histamine H1 Receptor(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
• [Presentation] G 蛋白共役受容体(GPCR)の結晶構造解析に向けた試み(シンポジウム)
  • Author(s): 小林拓也
  • Organizer: 第84回日本薬理学会年会
  • Place of Presentation: 横浜
• [Presentation] Crystallization of human membrane proteins using antibody fragments.
  • Author(s): Kobayashi T.
  • Organizer: Protein Island Matsuyama (PIM) International Symposium 2013
  • Place of Presentation: Matsuyama
  • Invited
• [Presentation] Crystallization of mammalian membrane proteins using antibody fragments.
  • Author(s): Kobayashi T.
  • Organizer: 4th International Symposium on Diffraction Structural Biology
  • Place of Presentation: Nagoya
  • Invited
• [Presentation] 機能性抗体を利用したGPCRのX線結晶構造解析への試み
  • Author(s): 小林拓也
  • Organizer: 千里ライフサイエンスセミナー
  • Place of Presentation: 大阪市
  • Invited
• [Presentation] GPCRのシグナル伝達分子機構の解明から創薬に向けて
  • Author(s): 小林拓也
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜市
  • Invited
• [Presentation] GPCRのX線結晶構造解析と創薬に必要なGPCRの構造情報について
  • Author(s): 小林拓也
  • Organizer: よこはまNMR構造生物学研究会 第47回ワークショップ
  • Place of Presentation: 横浜市
  • Invited
• [Presentation] GPCRをターゲットにした『構造に指南された創薬』を目指すための三つの戦略
  • Author(s): 小林拓也
  • Organizer: 構造活性フォーラム2012
  • Place of Presentation: 京都市
  • Invited
• [Presentation] GPCRをターゲットにしたX線結晶構造解析の現状と今後の展望
  • Author(s): 小林拓也
  • Organizer: 日本神経化学会公開シンポジウム
  • Place of Presentation: 神戸市
  • Invited
• [Presentation] GPCRの構造解析と抗体の利用
  • Author(s): 小林拓也
  • Organizer: 千里ライフサイエンスセミナー
  • Place of Presentation: 大阪市
• [Book] GPCRの立体構造から解き明かす生命科学 (2014)
  • Author(s): 小林拓也, 岩田想
  • Publisher: 羊土社
• [Book] G蛋白質共役受容体のX線結晶構造解析の進歩, 血栓と循環 (2013)
  • Author(s): 小林拓也
  • Publisher: メディカルレビュー社
• [Book] 「膜タンパク質」の重要性とその構造が拓く未来, 膜タンパク質構造研究最前線 (2013)
  • Author(s): 村田武士, 小林拓也, 野村紀通, 岩田想
  • Publisher: 化学同人
• [Book] ムスカリン性アセチルコリン受容体の構造解析から創薬に向けて, 膜タンパク質構造研究最前線 (2013)
  • Author(s): 小林拓也
  • Publisher: 化学同人
• [Book] 「膜タンパク質」の重要性とその構造が拓く未来 膜タンパク質構造研究 最前線 (2013)
  • Author(s): 村田武士、小林拓也、野村紀通、岩田想
  • Publisher: 化学同人
• [Book] ムスカリン性アセチルコリン受容体の構造解析から創薬に向けて 膜タンパク質構造研究 最前線 (2013)
  • Author(s): 小林拓也
  • Publisher: 化学同人
• [Book] 膜タンパク質の発現と精製 「昆虫細胞による生産」 膜タンパク質構造研究 最前線 (2013)
  • Author(s): 浅田秀基、小林拓也
  • Publisher: 化学同人
• [Book] GPCRのX線結晶構造解析に成功するための技術的進展, 実験医学 (2012)
  • Author(s): 小林拓也, 岩田想
  • Publisher: 羊土社
• [Book] ムスカリン受容体の立体構造が明らかに, 医学のあゆみ (2012)
  • Author(s): 小林拓也
  • Publisher: 医歯薬出版
• [Book] Gタンパク共役受容体(GPCR)の三次元結晶構造解析への試み, バイオインダストリー (2011)
  • Author(s): 小林拓也
  • Publisher: シーエムシー出版
• [Remarks]
  • URL: http://cell.mfour.med.kyoto-u.ac.jp/
• [Remarks] http://cell.mfour.med.kyoto-u.ac.jp/
• [Patent(Industrial Property Rights)] アゴニストの親和性を亢進する抗ヒトアデノシンA2a受容体モノクローナル抗体 (2012)
  • Inventor(s): 小笠原諭, 日野智也, 島村達郎 , 荒川孝俊, 万木貴美, 寿野千代, 村田武士, 小林拓也, 岩田想
  • Industrial Property Rights Holder: 独立行政法人科学技術振興機構
  • Industrial Property Rights Type: 特許
  • Filing Date: 2012-03-06
• [Patent(Industrial Property Rights)] 抗ヒトバンド3モノクローナル抗体 (2011)
  • Inventor(s): 小林拓也, 小林貴美, 荒川孝俊, 日野智也, 村田武士, 野村紀通, 岩田想, 浜窪隆雄, 岩成宏子, 望月康弘, 濱崎直孝
  • Industrial Property Rights Holder: 独立行政法人科学技術振興機構
  • Industrial Property Rights Type: 特許
  • Filing Date: 2011-11-10