Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry

Abstract Understanding the effect of temperature to the structural integrity of proteins is relevant to diverse areas such as biotechnology and climate change. Variable temperature ion mobility-mass spectrometry (VT-IM-MS) can measure the effect of temperature on conformational landscapes. To deline...

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Bibliographic Details
Main Authors: Xudong Wang, Emma Norgate, Junxiao Dai, Florian Benoit, Tony Bristow, Richard M. England, Jason M. D. Kalapothakis, Perdita E. Barran
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59065-x
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Summary:Abstract Understanding the effect of temperature to the structural integrity of proteins is relevant to diverse areas such as biotechnology and climate change. Variable temperature ion mobility-mass spectrometry (VT-IM-MS) can measure the effect of temperature on conformational landscapes. To delineate collision effects from structural change we report measurements using molecules with different degrees of rigidity namely: poly (L-lysine) (PLL) dendrimer, ubiquitin, β-casein and α-synuclein from 190-350 K. The CCS of PLL dendrimer varies with temperature consistent with collision theory, by contrast, the structure of each protein alters with notable restructuring at 350 K and 250 K, following predicted in vitro stability curves. At 210 K and 190 K we kinetically trap unfolding intermediates. For alpha-synuclein, the 13+ ions present two distinct conformers and VT-IM-MS measurements allow us to calculate the transition rate and activation energies of their conversion. These data exemplify the capacity of VT-IM-MS to provide insights on the thermodynamics of conformational restructuring.
ISSN:2041-1723