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


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.

SUBMITTER: Gessner C 

PROVIDER: S-EPMC5476592 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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

Gessner Chris C   Steinchen Wieland W   Bédard Sabrina S   J Skinner John J   Woods Virgil L VL   Walsh Thomas J TJ   Bange Gert G   Pantazatos Dionysios P DP  

Scientific reports 20170619 1


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. H  ...[more]

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