Scientific Reports (Jun 2017)

Computational method allowing Hydrogen-Deuterium Exchange Mass Spectrometry at single amide Resolution

  • Chris Gessner,
  • Wieland Steinchen,
  • Sabrina Bédard,
  • John J. Skinner,
  • Virgil L. Woods,
  • Thomas J. Walsh,
  • Gert Bange,
  • Dionysios P. Pantazatos

DOI
https://doi.org/10.1038/s41598-017-03922-3
Journal volume & issue
Vol. 7, no. 1
pp. 1 – 10

Abstract

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Abstract Hydrogen-deuterium exchange (HDX) coupled with mass spectrometry (HDXMS) is a rapid and effective method for localizing and determining protein stability and dynamics. Localization is routinely limited to a peptide resolution of 5 to 20 amino acid residues. HDXMS data can contain information beyond that needed for defining protein stability at single amide resolution. Here we present a method for extracting this information from an HDX dataset to generate a HDXMS protein stability fingerprint. High resolution (HR)-HDXMS was applied to the analysis of a model protein of a spectrin tandem repeat that exemplified an intuitive stability profile based on the linkage of two triple helical repeats connected by a helical linker. The fingerprint recapitulated expected stability maximums and minimums with interesting structural features that corroborate proposed mechanisms of spectrin flexibility and elasticity. HR-HDXMS provides the unprecedented ability to accurately assess protein stability at the resolution of a single amino acid. The determination of HDX stability fingerprints may be broadly applicable in many applications for understanding protein structure and function as well as protein ligand interactions.