| Project/Area Number |
24241062
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Genome biology
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
SUGIYAMA Naoyuki 京都大学, 大学院薬学研究科, 准教授 (50545704)
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| Co-Investigator(Renkei-kenkyūsha) |
TOYOSHIMA Fumiko 京都大学, ウイルス研究所, 教授 (40397576)
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 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥47,320,000 (Direct Cost: ¥36,400,000、Indirect Cost: ¥10,920,000)
Fiscal Year 2014: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
Fiscal Year 2013: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2012: ¥20,150,000 (Direct Cost: ¥15,500,000、Indirect Cost: ¥4,650,000)
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| Keywords | シグナル伝達 / プロテオーム / リン酸化 / キナーゼ |
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| Outline of Final Research Achievements |
In this study, we have successfully developed the analytical system to acquire the high-precision kinase substrate motifs based on a large-scale in vitro kinase assay as well as substrate prediction scoring methods. Moreover, we further developed a workflow for quantitative phosphoproteomics using meter-scale monolithic silica capillary columns combined with multiplex quantitation tags, and applied to phosphorylation perturbation studies. As a result, this system allowed to identify intracellular kinases as well as their substrates regulated by various perturbations, by converting the phosphoproteome fluctuations to kinase variation. Since the throughput of the measurement system is practical (1 hour per analysis), this system could contribute to the evolution of the molecular basis of intracellular phosphorylation network by acquiring a more extensive data in future.
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