Use of highly efficient procedure in getting revertant cells, for studies of interaction manner between membrane protein and its inhibitor
| Project/Area Number |
17K08274
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | The University of Tokushima |
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Principal Investigator |
SHINOHARA Yasuo 徳島大学, 先端酵素学研究所(プロテオ), 教授 (60226157)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | ADP/ATP輸送体 / ボンクレキン酸 / 分子間相互作用 / 復帰変異株 / タンパク質リガンド相互作用 / ミトコンドリア |
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| Outline of Final Research Achievements |
We tried to isolate the revertant yeast cells viable on the glycerol plate containing bongkrekic acid, by screening the library of yeast cells expressing randomly mutated mitochondrial ADP/ATP carrier. As a result, we succeeded in identifying two known mutations of L142S and G298S, and three unknown mutations of I200V, S245P and V300I. Furthermore, these identified amino acids were shown to locate closely to bongkrekic acid, by independently achieved crystallographic analysis of the ADP/ATP carrier complexed with bongkrekic acid.
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| Academic Significance and Societal Importance of the Research Achievements |
膜タンパク質とリガンドの相互作用の様式の理解は、創薬を考える上で極めて大きな課題であるが、これまでは光親和性標識かタンパク質のX線結晶構造解析しか実験法が確立されておらず、新たな研究手法の開発が急務であった。
今回の研究によって、我々が見出した高効率な復帰変異株獲得による膜蛋白質とリガンドの相互作用解析法が実用的なものであることが示され、新たな研究手法になり得るものと期待される。
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Report
(4 results)
Research Products
(15 results)
