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Updated: Jul 8, 2026

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
Published on: November 29, 2013
Hydrogen Exchange Mass Spectrometry (HX-MS) for Mapping Antibody Binding Sites and Conformational Dynamics of
Vikram G Pillai1, X Long Zheng2,3
1Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS, USA.
Abstract:
Hydrogen exchange mass spectrometry (HX-MS) enables the determination of subtle conformational and dynamic changes in proteins under near-physiological conditions. Exchange of backbone amide hydrogens in proteins with deuterium occurs in solution and is influenced by solvent accessibility, hydrogen bonding, and structural dynamics. In this protocol, we employ HX-MS to investigate conformational dynamics and map antibody binding sites of ADAMTS13 using the N-terminal truncated variant MDTCS (e.g., metalloprotease, disintegrin, thrombospondin type 1 repeat, cysteine-rich, and spacer domain). Deuterium incorporation is monitored over multiple time points and mapped onto known protein structures to reveal protected and flexible regions, which infers the antibody binding epitope on ADAMTS13. This method is applicable to study any other proteins that interact with their ligand or antibodies.
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