Development of the strategy for creating stabilized GPCR variants for large-scale preparation
| Project/Area Number |
25709080
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| Research Category |
Grant-in-Aid for Young Scientists (A)
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| Allocation Type | Partial Multi-year Fund |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Kyushu University |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥20,800,000 (Direct Cost: ¥16,000,000、Indirect Cost: ¥4,800,000)
Fiscal Year 2016: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2015: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2013: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
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| Keywords | G蛋白質共役型受容体 / 出芽酵母 / 安定化 / 大量調製 / 膜蛋白質 / 生物機能工学 / 蛋白質工学 |
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| Outline of Final Research Achievements |
The aim of this project is to develop the strategy for stabilization of unstable G-protein coupled receptors (GPCRs) for large-scale preparation. We tried to establish the screening system of stabilized GPCRs using the signal transduction system in S. cerevisiae. We also established the rapid screening system in 96-well plate format. Using this system, stabilized variants of GPCR_A, whose structure has not solved, were successfully selected. Then we established the large-scale expression strategy in S. cerevisiae. The stabilized variants of GPCR_A were purified in milligram quantities, and the ligand-receptor interactions were biophysically examined. We also tried crystallization of the purified GPCR_A for determination of its structure.
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Report
(5 results)
Research Products
(24 results)
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[Journal Article] Proteoliposome-based selection of a recombinant antibody fragment against the human M2 muscarinic acetylcholine receptor.2014
Author(s)
Suharni, Nomura Y, Arakawa T, Hino T, Abe H, Nakada-Nakura Y, Sato Y, Iwanari H, Shiroishi M, Asada H, Shimamura T, Murata T, Kobayashi T, Hamakubo T, Iwata S, Nomura N.
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Journal Title
Monoclon Antib Immunodiagn Immunother
Volume: 33
Issue: 6
Pages: 378-385
DOI
NAID
Related Report
Peer Reviewed
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