High-Throughput Determination of Exchange Rates of Unmodified and PTM-Containing Peptides Using HX-MS.

ABSTRACT: Despite the widespread use of MS for hydrogen/deuterium exchange measurements, no systematic, large-scale study has been conducted to compare the observed exchange rates in protein-derived, unstructured peptides measured by MS to the predicted exchange rates calculated from NMR-derived values and how neighboring residues and post-translational modifications influence those exchange rates. In this study, we sought to test the accuracy of predicted values by performing hydrogen exchange measurements on whole cell digests to generate an unbiased dataset of 563 unique peptides derived from naturally occurring protein sequences. A remarkable 97% of observed exchange rates of peptides are within two-fold of predicted values. Using fully deuterated controls, we found that for approximately 50% of

SUBMITTER: Moroco JA 

PROVIDER: S-EPMC11875167 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature