Elucidation of intracellular phosphorylation network for molecular targeting drug discovery

2014 Fiscal Year Final Research Report

• Project/Area Number: 24241062
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Genome biology
• Research Institution: Kyoto University
• Principal Investigator: ISHIHAMA Yasushi 京都大学, 薬学研究科(研究院), 教授 (30439244)
• Co-Investigator(Kenkyū-buntansha): SUGIYAMA Naoyuki 京都大学, 大学院薬学研究科, 准教授 (50545704)
• 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)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: シグナル伝達 / プロテオーム / リン酸化 / キナーゼ

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.

Free Research Field

プロテオミクス