Purification and functional analysis of YhaTU system, a two component K+/H+ antiporter from Bacillus subtilis
| Project/Area Number |
23770156
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Functional biochemistry
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| Research Institution | Toyo University |
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Principal Investigator |
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| Project Period (FY) |
2011 – 2013
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | イオンチャネル / トランスポーター / アンチポーター / チャネル / 部位特異的変異 / タンパク質精製 / K+/H+アンチポーター / 枯草菌 |
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| Research Abstract |
Bacillus subtilis YhaTU system is a K+/H+ antiporter which consists from two components, the hydrophilic ancillary protein YhaT and the membrane-embedded YhaU. The YhaTU system is not only a unique cation proton antiporter but also an interesting enzyme in respect of the evolution of ion antiporter from ion channel. In In this study, purification of the YhaTU system and cloning the genes which have point mutations at the various points of the genes have been tried.
To purify the YhaTU complex, StrepTagII and His6 tags were introduced at N-terminal or C-terminal of the proteins. Fifty six mutations were also introduced into the genes to test the function of each amino acid residues. Now the basis to analyze the detail function of YhaTU has been constructed.
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Report
(4 results)
Research Products
(10 results)
