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Updated: Jun 13, 2026

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
Published on: April 17, 2017
Combination of single-molecule Förster resonance energy transfer and hydrogen-deuterium exchange mass spectrometry
Wei Chen1, Elizabeth A Komives2
1Department of Chemistry, The Pennsylvania State University, University Park, PA, USA; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
Abstract:
Understanding how proteins function requires a description of their structural dynamics. Accessing these structural dynamics across time and length scales calls for different methods and often their combination. Here we review a combination of two highly synergistic and complementary methods-single-molecule Förster resonance energy transfer (smFRET) and hydrogen-deuterium exchange mass spectrometry (HDX-MS). We describe how they complement each other in the ability to resolve structural and dynamical heterogeneity, coverage of protein residues, and types of protein motions they probe. Together, they access a wide range of protein dynamics from global rigid body motions to local unfolding. This synergy has led to the elucidation of activation and regulatory mechanisms in signaling and transcription, which we review in five case studies, highlighting the key role these two methods play in advancing dynamic structural biology.
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