| 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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| 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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