Hydrogen exchange mass spectrometry for studying protein structure and dynamics

Area of Science:

• Biochemistry
• Structural Biology
• Analytical Chemistry

Background:

• Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is crucial for studying protein structure and dynamics in solution.
• HDX-MS leverages the differential exchange rates of amide hydrogens with deuterium in disordered versus structured protein regions.

Purpose of the Study:

• To provide a tutorial review of HDX-MS fundamentals.
• To highlight recent advancements and applications in HDX-MS.
• To serve as a reference for both novices and practitioners in the field.

Main Methods:

• Utilizes mass spectrometry (MS) for peptide mapping to measure mass shifts.
• Explores gas phase fragmentation strategies as alternatives to traditional proteolysis.
• Discusses electron-based dissociation methods to overcome scrambling phenomena.

Main Results:

• Recent developments enhance HDX-MS applicability to membrane proteins.
• Advances facilitate the characterization of short-lived protein folding intermediates.
• Gas phase fragmentation offers a promising avenue, with electron dissociation mitigating scrambling.

Conclusions:

• HDX-MS is a powerful tool for protein structural and dynamic analysis.
• Emerging fragmentation techniques and applications expand the utility of HDX-MS.
• This review offers a comprehensive overview of current trends in HDX-MS.