Conformation change of synaptotagmin probed by the fluorescent unnatural amino acid in vivo
| Project/Area Number |
16K07072
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurochemistry/Neuropharmacology
|
|---|
| Research Institution | Osaka Medical College |
|---|
Principal Investigator |
Sakata Souhei 大阪医科大学, 医学部, 准教授 (40528006)
|
|---|
| Project Period (FY) |
2016-04-01 – 2020-03-31
|
| Project Status |
Completed (Fiscal Year 2019)
|
| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
|
|---|
| Keywords | シナプス / シナプトタグミン / 遺伝コード拡張法 / 非天然アミノ酸 / イオンチャネル / ゼブラフィッシュ / 立体構造変化 / アセチルコリン受容体 / 遺伝コード拡張 / シナプス前終末 / 神経筋接合部 / 膜融合 / タンパク質の立体構造変化 |
|---|
| Outline of Final Research Achievements |
Synaptotagmin (Syt) is known to be the calcium sensor in presynaptic terminals. I have carried out my project to examine conformation changes of Syt in vivo using the genetic code expansion method. This method enables to incorporate the fluorescent unnatural amino acid (Anap) in the polypeptide of target protein as a fluorescent marker for conformation changes. First, I established the technique to apply the method in zebrafish embryo. Second, I searched the position into which Anap is incorporated using acethylcholine receptors and showed that only one site is available for the measurement of Anap fluorescence change. This means that limited positions can be used as an incorporation site to probe conformation change. These are big steps for the measurement of protein structure changes in living animals.
|
| Academic Significance and Societal Importance of the Research Achievements |
遺伝コード拡張法はタンパク質の狙った部位特異的に蛍光タグを導入できるなど、非常に有用な方法ではあるが、培養細胞等や一部の発現系などにしか適用できなかった。本研究課題ではこれを生体に応用することに成功した。これは生体で遺伝コード拡張法を機能させた数少ない例である。また今後、他のタンパク質でも同様に細胞内での構造変化に関する情報が得られれば、新規の治療薬の開発につながる可能性もある。
|
Report
(5 results)
Research Products
(10 results)
