Technologicaldevelopment for construction of synthetic proteins that facilitate membrane protein crystallization
Project/Area Number |
22570114
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Structural biochemistry
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Research Institution | Kyoto University |
Principal Investigator |
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Co-Investigator(Renkei-kenkyūsha) |
IWATA So 京都大学, 大学院・医学研究科, 教授 (60452330)
|
Project Period (FY) |
2010 – 2012
|
Project Status |
Completed (Fiscal Year 2012)
|
Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
|
Keywords | 膜蛋白質 / X線結晶構造解析 / 結晶化リガンド / 抗体フラグメント / ファージディスプレイ / 膜輸送体 / プァージディスプレイ |
Research Abstract |
Co-crystallization of membrane proteins with antibody fragments may emerge as a general tool to facilitate crystal growth and improve crystal quality. The bound antibody fragment enlarges the hydrophilic part of the mostly hydrophobic membrane protein, thereby increasing the interaction area for possible protein-protein contacts in the crystal. To serve as a ‘crystallizing ligand’, the antibody fragment has to recognize an epitope that is only present in the native conformation (and not in the denatured state) of the membrane protein, bind with high affinity, and form stable and rigid complexes. In this project, we developed a fast and reliable method for generating crystallizing ligands against various mammalian G-protein coupled receptors (GPCRs) and transporters in a phage display format. Using this method, the crystal structure of a mammalian facilitative glucose transporter complexed with a Fv fragment has been determined at 3.5Åresolution.
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Report
(4 results)
Research Products
(10 results)
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[Journal Article] Cloning, expression and purification of the anion exchanger 1 homologue from the basidiomycete Phanerochaete chrysosporium2011
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.
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Journal Title
Protein. Expr. Purif.
Volume: 79(1)
Issue: 1
Pages: 81-7
DOI
NAID
Related Report
Peer Reviewed
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[Journal Article] Evaluation of the Pichia pastoris expression system for the production of GPCRs for structural analysis2011
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
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Journal Title
Microbial Cell Factories
Volume: 10
Issue: 1
Pages: 24-24
DOI
Related Report
Peer Reviewed
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[Patent(Industrial Property Rights)] 膜蛋白質の立体構造を認識するモノクローナル抗体のスクリーニング方法2010
Inventor(s)
日野智也, 村田武士, 荒川孝俊, 野村紀通, 小林拓也, 岩田想
Industrial Property Rights Holder
日野智也, 村田武士, 荒川孝俊, 野村紀通, 小林拓也, 岩田想
Filing Date
2010-04-28
Related Report
Overseas