Dynamics of hydrogen bond network among the side chains in the active site of enzyme revealed by NMR deuterium isotope shift

2015 Fiscal Year Final Research Report

• Project/Area Number: 26650023
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Hiroshima University
• Principal Investigator: Shin-ichi Tate 広島大学, 理学(系)研究科(研究院), 教授 (20216998)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: NMR / プロリン異性化酵素 / 重水素同位体シフト / 安定同位体 / タンパク質 / 酵素

Outline of Final Research Achievements

NMR isotope shifts caused by the proton/deuteron change were exploited to reveal the relations between the proline isomerase activity of Pin1 and the hydrogen bond network formed in the active site. The isotope shift was used to quantitatively elucidate the changes in the strength of the hydrogen bonds in the network. We made three different mutants having amino acid changes to C113 in the active site of the isomerase domain of Pin1, PPIase. In addition, C138 mutation was applied to see the distal effect on the hydrogen bond network.
Combined the results from the analyses on the different mutants made the role of the hydrogen bond network in the PPIase active site clearer.

Free Research Field

生物物理化学