Multiple analysis of FMN-binding reaction and consideration on molecular evolution based on structural information

2016 Fiscal Year Final Research Report

• Project/Area Number: 26450131
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Osaka City University
• Principal Investigator: KITAMURA Masaya 大阪市立大学, 大学院工学研究科, 教授 (20244634)
• Research Collaborator: TSUMOTO Kouhei
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: フラボタンパク質 / タンパク質工学 / 構造機能相関 / 補因子 / 国際情報交換

Outline of Final Research Achievements

Binding reaction analysis with cofactor in FMN-binding protein (FMN-bp) was performed. Binding strength with cofactor was thermodynamically analyzed by the surface plasmon resonance spectroscopy and isotherm titration calorimetry in addition to the fluorescence spectroscopy. The dissociation constant with cofactor was not affected by the phosphate ion concentration. By changing the preparation method of apoprotein, I could obtain a large amount of apoFMN-bp having binding activity, and determined the three-dimensional structure. These results were compared with other flavoproteins and new proposals were suggested on the binding mechanism of apoFMN-bp and FMN.

Free Research Field

分子生物学