Real-time tracking of protein unfolding with time-resolved x-ray solution scattering

L. Henry, M. R. Panman, L. Isaksson, E. Claesson, I. Kosheleva, R. Henning, S. Westenhoff, O. Berntsson

Research output: Contribution to journalArticlepeer-review

3 Citations (SciVal)

Abstract

The correct folding of proteins is of paramount importance for their function, and protein misfolding is believed to be the primary cause of a wide range of diseases. Protein folding has been investigated with time-averaged methods and time-resolved spectroscopy, but observing the structural dynamics of the unfolding process in real-time is challenging. Here, we demonstrate an approach to directly reveal the structural changes in the unfolding reaction. We use nano- to millisecond time-resolved x-ray solution scattering to probe the unfolding of apomyoglobin. The unfolding reaction was triggered using a temperature jump, which was induced by a nanosecond laser pulse. We demonstrate a new strategy to interpret time-resolved x-ray solution scattering data, which evaluates ensembles of structures obtained from molecular dynamics simulations. We find that apomyoglobin passes three states when unfolding, which we characterize as native, molten globule, and unfolded. The molten globule dominates the population under the conditions investigated herein, whereas native and unfolded structures primarily contribute before the laser jump and 30 μs after it, respectively. The molten globule retains much of the native structure but shows a dynamic pattern of inter-residue contacts. Our study demonstrates a new strategy to directly observe structural changes over the cause of the unfolding reaction, providing time- and spatially resolved atomic details of the folding mechanism of globular proteins.

Original languageEnglish
Article number054702
JournalStructural Dynamics
Volume7
Issue number5
DOIs
Publication statusPublished - 2020

Subject classification (UKÄ)

  • Biochemistry and Molecular Biology

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