Presentation Information
[P34]NMR analysis of transient unfolding/refolding protein structures by encapsulation in a self-assembled cage
*Anouk Rossen1, Takahiro Nakama1, Maho Yagi-Utsumi2, Daishi Fujita3, Koichi Kato2, Makoto Fujita1 (1. Graduate School of Engineering, The University of Tokyo, 2. Institute for Molecular Science (IMS), 3. Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University)
Keywords:
Protein,Unfolding,Refolding,Self-assembled cage,Transient structures
We analysed transient protein structures in solvent-induced unfolding and refolding by encapsulation of a protein in a self-assembled cage. The confined space in the cage can isolate an unstable unfolded protein, preventing formation of insoluble aggregates, enabling NMR analysis. 1H–15N HSQC NMR indicated a transition to a transient unfolded intermediate by sudden peak attenuation with increasing acetonitrile solvent. Mapping of peak intensity reduction revealed perturbations concentrated in certain regions, suggesting that collapse of those regions initiated the unfolding. By decreasing the acetonitrile ratio, we also examined the refolding of the denatured protein, revealing the transient structures of unfolding/refolding processes.