| Project/Area Number |
25410139
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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 |
Analytical chemistry
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| Research Institution | Saitama University |
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Principal Investigator |
SAITO Shingo 埼玉大学, 理工学研究科, 准教授 (60343018)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2015: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2013: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
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| Keywords | メタロミクス / 金属タンパク質 / 電気泳動 / スペシエーション / ゲル電気泳動 |
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| Outline of Final Research Achievements |
Although investigation of metalloproteins is of importance to realize the relationship between metal ion and biological behavior, identification of metalloproteins is difficult. Here, we have developed three types of new polyacrylamide gel electrophoresis (PAGE) specific to metalloproteins and protein-bound metal ions in biological samples: metal ion contaminant sweeping (MICS)-Blue Native (BN)-PAGE; metal ion detection PAGE; and holo-apo conversion two dimensional MICS-PAGE.
In MICS-BN-PAGE, all metal contaminants were excluded from the gel, the separation buffer and the instruments, resulting in protein separation under metal-free conditions. In metal detection PAGE, metal ions (Fe, Cu and others) were separated and fluorescently detected with high sensitivity (to the ppt level) by the addition of probes. In holo-apo conversion 2D MICS-BN-PAGE, metalloproteins were exclusively isolated off of the diagonal line.
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