Creation of the analysis technique to visualize the dynamics of labile intermediates in phosphotransfer reactions

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K07887
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: Hiroshima University
• Principal Investigator: Kinoshita Emiko 広島大学, 医歯薬保健学研究科(薬), 助教 (40379912)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: phos-tag / チオリン酸化タンパク質 / ATPgammaS

Outline of Final Research Achievements

Signal-transduction regulatory systems for certain cellular responses which rely on phosphorylation of histidine or aspartic acid have known in eukaryotic cells. Identification of site-specific phosphorylation dynamics of His or Asp is technically difficult because of the labile nature of the phosphorylated amino acid residues. ATPgammaS, which used as a phosphoryl donor to trace protein kinase activities, has the advantage of permitting study of His- or Asp phosphorylation, because the resulting thiophosphorylated substrates are resistant to chemical hydrolysis or the action of phosphatases. Using the phosphate affinity electrophoresis, Phos-tag SDS-PAGE, we separated thiophosphorylated substrate generated as a intermediate of kinase reaction，and studied the dynamics of the histidine kinase reactions.

Free Research Field

物理系薬学