Crystallization of membrane proteins by combined application of LCP and LSP methods

• Project/Area Number: 25650019
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: The University of Tokyo
• Principal Investigator: NISHIZAWA Tomohiro 東京大学, 理学(系)研究科(研究院), 助教 (80599077)
• Project Period (FY): 2013-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2013: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
• Keywords: X線結晶構造解析 / 膜タンパク質 / 結晶化

Research Abstract

We developed a crystallization method for membrane proteins. Generally, membrane proteins are not stable in detergent micelles, disturbing the crystallization process. Recently Lipidic Cubic Phase (LCP) methods have been applied to obtain membrane protein crystals in lipid-like environment, which advances structural studies of membrane proteins. However its application range is limited to GPCRs and only a small number of other transporters. We found that LCP and its modified method LSP can be applied in combination to facilitate membrane protein crystallization and the application range is not limited. These techniques have lead to the successful crystallization of Ca2+/H+ exchanger (CAX), tight junction forming protein (claudin), and other membrane proteins.

Research Products

• [Journal Article] Crystal structure of a claudin provides insight into the architecture of tight junctions (2014)
  • Author(s): Suzuki H, Nishizawa T, Tani K, Yamazaki Y, Tamura A, Ishitani R, Dohmae N, Tsukita S, Nureki O, Fujiyoshi Y
  • Journal Title: Science Volume: 344 Issue: 6181 Pages: 304-307
  • DOI: 10.1126/science.1248571
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Crystal structure of a claudin provides insight into the architecture of tight junctions (2013)
  • Author(s): Suzuki H、Nishizawa T、 Tani K、Yamazaki Y、Tamura A、 Ishitani R、Dohmae N、Tsukita S、 Nureki O、Fujiyoshi Y
  • Journal Title: Science Volume: 344巻 Pages: 304-307
  • Peer Reviewed
• [Journal Article] Structural basis for the counter-transport mechanism of a H+/Ca2+ exchanger (2013)
  • Author(s): Nishizawa T、Kita S、Maturana AD、Furuya N、Hirata K、 Kasuya G、Ogasawara S、Dohmae N、 Iwamoto T、Ishitani R、Nureki O
  • Journal Title: Science Volume: 341巻 Pages: 168-172
  • Peer Reviewed
• [Journal Article] Structural Basis for the Counter-Transport Mechanism of a H+/Ca2+ Exchanger (2013)
  • Author(s): Nishizawa T, Kita S, Maturana AD, Furuya N, Hirata K, Kasuya G, Ogasawara S, Dohmae N, Iwamoto T, Ishitani R, Nureki O
  • Journal Title: Science Volume: 341 Issue: 6142 Pages: 168-172
  • DOI: 10.1126/science.1239002
  • Peer Reviewed
• [Presentation] LCP結晶化ノウハウ (2013)
  • Author(s): 西澤知宏
  • Organizer: 平成25年度日本結晶学会年会および総会
  • Place of Presentation: 熊本大学(熊本)
  • Year and Date: 2013-10-13
  • Invited
• [Presentation] プロトン・カルシウム交換輸送体における構造基盤 (2013)
  • Author(s): 西澤知宏、石谷隆一郎、古屋憲孝、喜多紗斗美、岩本隆宏、アンドレアスマツラナ、濡木理
  • Organizer: 第86回日本生化学大会インターナショナルシンポジウム
  • Place of Presentation: パシフィコ横浜(神奈川)
  • Year and Date: 2013-09-13
  • Invited
• [Presentation] LCP結晶化ノウハウ (2013)
  • Author(s): 西澤知宏
  • Organizer: 平成25年度 日本結晶学会 年会および総会
  • Place of Presentation: 熊本大学(熊本）
  • Invited
• [Presentation] Structural Basis for the Counter-transport Mechanism of a H+/Ca2+ Exchanger (2013)
  • Author(s): 西澤知宏, 石谷隆一郎, 古屋憲孝, 喜多紗斗美､ 岩本隆宏, アンドレアス マツラナ, 濡木理
  • Organizer: 第86回日本生化学大会インターナショナルシンポジウム
  • Place of Presentation: 横浜パシフィコ（神奈川）
  • Invited