| Project/Area Number |
18570070
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Animal physiology/Animal behavior
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| Research Institution | Hiroshima University |
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Principal Investigator |
UEKI Tatsuya Hiroshima University, Graduate School of Science, Associate Professor (10274705)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,050,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥450,000)
Fiscal Year 2007: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2006: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | ascidians / vanadium / redox / metal accumulation / micro array / タンパク質 / グルタチオン / システイン / 金属結合タンパク質 |
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| Research Abstract |
The purpose of this study is to reveal the metal binding domains of a vanadium-binding protein Vanabin2, to reveal the relationship between disulfide banding structures and metal binding, and to reveal the redox reaction and biological function of vanadium in ascidians.
1. Several mutant vanabins were constructed and their metal binding ablitily was assayed. lysine residues located near the 64th histidine are important for high affinity binding of vanadium(IV).
2. Serine substitutions of cysteines caused structural instability and degradation of the mutant proteins. Especially fifth disulfide bonding was most important for structural stability These mutation did not affect the binding of vanadium.
3. Metal selectivity of glutathione transferase (AsGST) was assayed. AtpH4.5, AsGST bound specifically to iron (III), copper(II) and vanadium (IV).
4. When Vanabin2 was treated with several reductants in vitro, intermediate structures were observed. Relationship between the reduction of vanadium(V) and the redox of disulfide bonds in Vanabin2 was assessed in vitro. Redox intermediate structure of Vanabin2 was suggested.
5. By micro array analysis on Ciona intestinalis, genes whose expression was affected by the addition of vanadium(IV) and vanadium(V) ions were explored. Basic sugar metabolism pathways were not affected, except for pentose phosphate pathway. In addition, enzymes in glutathione metabolism were affected.
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