| Project/Area Number |
24659017
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Physical pharmacy
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
WAKABAYASHI Masaki 京都大学, 大学院薬学研究科, 助教 (70552024)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2013: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2012: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
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| Keywords | プロテオーム / 血漿プロテオーム / 分析科学 / 蛋白質 / 血清プロテオーム |
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| Research Abstract |
We aimed to develop a novel method to enrich low abundant proteins, in which target proteins are labeled with phosphate by in vitro phosphorylation reaction using selected spiked kinases, followed by phosphopeptide enrichment, for applying to plasma/serum proteome analysis. As a result, by combining the in vitro phosphorylation by optimal kinase with sample treatment by antibody-based serum depletion column, it becomes possible to identify several cytokines with very low expression level in plasma, which have never been reported to identify from plasma by conventional proteomics approaches due to their low abundance. Furthermore, we successfully quantified low abundant phosphorylated proteins in cultured cells by this method, which were not possible by the conventional methods.
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