Establishment of a quantitative method for the dynamical analysis of bacterial two-component systems toward studies on an innovative drug development

2015 Fiscal Year Final Research Report

• Project/Area Number: 25560417
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Chemical biology
• Research Institution: Hiroshima University
• Principal Investigator: Kinoshita Eiji 広島大学, 医歯薬保健学研究院(薬), 准教授 (80304418)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: フォスタグ / ２成分伝達系 / ヒスチジンキナーゼ / レスポンスレギュレーター / 生体分子計測

Outline of Final Research Achievements

One mode of bacterial intracellular signal transduction involves two-component regulatory systems mediated by phosphorylation of His and Asp residues. The chemical instability of phosphate groups on these residues hampers research on protein phosphorylation in such regulatory systems. In this study, Phos-tag SDS-PAGE originally developed as a novel method to research on protein phosphorylation has been applied to the dynamical analysis of two-component systems. As a result, this method permitted to identify aspects relevant to signaling molecules, including kinetic properties of enzymes and substrates in intramolecular and intermolecular phosphotransfer and autophosphorylation reactions. Furthermore, Phos-tag SDS-PAGE has been demonstrated as a useful quantitative method for the development of inhibitors of histidine kinases.

Free Research Field

物理系薬学