| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Outline of Final Research Achievements |
There are some G protein-coupled receptors (GPCR) coupled to G12/13 protein, such as endothelin B receptor and angiotensin 1 receptor. However, the search of novel bioactive peptide based on the activation of G12/13 signaling pathway has not been successful so far, because it has been difficult to easily detect such activation.
The SRF-RE luciferase reporter is activated in response to activation of GPCR coupled to G12/13 protein. In this study, it was demonstrated that this reporter gene assay is suitable to search novel bioactive peptide that activates G12/13 signaling pathway through GPCR. This assay was detected that endothelin-1 induced dose-dependent, robust increase in SRF-RE reporter activity in HEK293 cells expressing endothelin B receptor with EC50 of 0.3nM, indicating that the sensitivity of this assay is sufficient for searching the novel bioactive peptide. Therefore, we are searching the novel bioactive peptide as endogenous ligand for orphan GPCRs by using this assay.
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