Challenge for heterodimer formation of endothelin receptors in nanodiscs
| Project/Area Number |
26640102
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Tumor therapeutics
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| Research Institution | Kyoto University |
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Principal Investigator |
Doi Tomoko 京都大学, 理学(系)研究科(研究院), 准教授 (00397580)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | 分子標的治療 / エンドセリン受容体 / ナノディスク / アンタゴニスト / アゴニスト / GPCR / 構造変化 |
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| Outline of Final Research Achievements |
In many cancer cells, expression levels of peptide hormone, endothelin and its receptors are elevated. However, molecular mechanisms of signal transduction and ligand binding by endothelin systems are not fully understood. The goals of this project are to reconstitute a heterodimer of endothelin receptor A-type (ETAR) and B-type (ETBR) in apoA-1 nanodiscs, and to compare its ligand binding properties with those of each homodimer in nanodiscs. The reconstitution system using ETBR well-expressed in insect cells, purified in detergent micelle solution, and apoA1 expressed in E.coli was established in this study. The ETBRs in nanodisc activate G proteins, Gi and Gq comparable to those reconstituted in phospholipid vesicles. However, efficient purification and reconstitution of ETAR in enough amounts could not be achieved and therefore formation of heterodimer was unsuccessful. Efforts to isolate ETAR in enough amounts by introducing overexpressing mutations will be continuously made.
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Report
(3 results)
Research Products
(2 results)
