High-energy conformations of proteins as a new target of structural biology

2015 Fiscal Year Final Research Report

• Project/Area Number: 25840025
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Ritsumeikan University
• Principal Investigator: Ryo Kitahara 立命館大学, 薬学部, 准教授 (70512284)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 圧力 / コンフォメーション / NMR / 熱力学

Outline of Final Research Achievements

The conventional NMR approach for structure determination so far reveals atomic coordinates of the most stable conformation of a protein, the so-called “native state”. Despite significant progress in our knowledge about the basic folded conformation of proteins, high-Gibbs free energy states of proteins are poorly understood. Here, we investigate the structure and backbone dynamics of locally disordered state of ubiquitin using high pressure NMR spectroscopy. We carried out NMR signal assignments, the collection of structural information based on chemical shifts, NOE, and paramagnetic relaxation enhancements (PRE). We also performed 15N spin relaxation experiments (T1, T2, hNOE). More than 1000 NOE- and PRE-based distance constraints and 42 torsion angle constraints were used for structure calculation of the locally disordered state. High-pressure NMR spectroscopy provides snapshots of fluctuating ubiquitin structure at atomic resolution.

Free Research Field

構造生物学