Development of a small-sized labeling method for membrane proteins and its application for heterooligomer analysis
| Project/Area Number |
18H02561
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Review Section |
Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
矢野 義明 京都大学, 薬学研究科, 講師 (60402799)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2020: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2019: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2018: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
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| Keywords | 膜タンパク質 / 蛍光イメージング |
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| Outline of Final Research Achievements |
To detect heterooligomer formation of membrane proteins, we developed novel tag-probe pairs that are orthogonal to our original E3-K4 coiled-coil labeling method. The EN3.5 tag-KN3.5 probe and EN24 tag-KN24 probe pairs containing one and two Asp residues, respectively, at the recognition site, were designed. The dissociation constants for these pairs were determined 48.7 and 20.4 nM, respectively, using β2AR. Furthermore, these pairs were orthogonal to the E3-K4 coiled-coil method, suggesting a possibility of detection of heterooligomers.
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| Academic Significance and Societal Importance of the Research Achievements |
細胞膜において異なる膜タンパク質同士が会合（ヘテロ会合）することが、膜タンパク質の機能発現に重要であり、異常なヘテロ会合は疾患の原因になることがある。しかし、生きた細胞上でこのヘテロ会合を精度良く検出する方法はこれまでになかった。本研究では、蛍光イメージングに用いることのできる膜タンパク質の新規標識法を開発し、我々がすでに開発した方法と併用することで、生きた細胞上で、膜タンパク質の機能を損なうことなく、精度良くヘテロ会合を検出する道を拓いた。
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Report
(4 results)
Research Products
(4 results)
