2016 Fiscal Year Final Research Report
Development of the strategy for creating stabilized GPCR variants for large-scale preparation
| Project/Area Number |
25709080
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (A)
|
| Allocation Type | Partial Multi-year Fund |
|---|
| Research Field |
Biofunction/Bioprocess
|
|---|
| Research Institution | Kyushu University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2013-04-01 – 2017-03-31
|
| 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 |
|---|
| 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.
|
| Free Research Field |
蛋白質工学
|
