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

LCP和LSP方法联合应用膜蛋白结晶

• 批准号: 25650019
• 负责人: NISHIZAWA Tomohiro
• 金额: $ 2.66万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2013
• 资助国家: 日本
• 起止时间: 2013-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-25650019/
• 关键词: X線結晶構造解析; 膜タンパク質; 結晶化

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.

我们开发了一种膜蛋白的结晶方法。通常，膜蛋白在洗涤剂胶束中不稳定，干扰结晶过程。近年来，脂质立方相（LCP）方法被用于在类脂质环境中获得膜蛋白晶体，推动了膜蛋白结构的研究。然而，其应用范围仅限于GPCR和少数其他转运体。我们发现，LCP及其改进的LSP方法可以联合应用，促进膜蛋白结晶，应用范围不受限制。这些技术已经导致Ca 2 +/H+交换器（CAX）、紧密连接形成蛋白（claudin）和其他膜蛋白的成功结晶。

项目成果

プロトン・カルシウム交換輸送体における構造基盤

质子和钙交换转运蛋白的结构基础

• 发表时间: 2013
• 作者: 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
• 通讯作者: 濡木理

LCP結晶化ノウハウ

LCP结晶技术

• 发表时间: 2013
• 作者: Kanoh;J.;西澤知宏
• 通讯作者: 西澤知宏

Crystal Structure of a Claudin Provides Insight into the Architecture of Tight Junctions

• DOI: 10.1126/science.1248571
• 发表时间: 2014-04-18
• 期刊: SCIENCE
• 影响因子: 56.9
• 作者: Suzuki, Hiroshi;Nishizawa, Tomohiro;Fujiyoshi, Yoshinori
• 通讯作者: Fujiyoshi, Yoshinori

Structural Basis for the Counter-Transport Mechanism of a H+/Ca2+ Exchanger

• DOI: 10.1126/science.1239002
• 发表时间: 2013-07-12
• 期刊: SCIENCE
• 影响因子: 56.9
• 作者: Nishizawa, Tomohiro;Kita, Satomi;Nureki, Osamu
• 通讯作者: Nureki, Osamu