Automated Specific Amino Acid Footprinting Mass Spectrometry: Repurposing an HDX Platform for Determining Reagent Feasibility.

ABSTRACT: Protein footprinting is a mass spectrometry (MS)-based approach to measure protein conformational changes. One approach, specific amino acid labeling, imparts often an irreversible modification to protein side chains but requires careful selection of the reactive reagent and often time-consuming optimization of experimental parameters prior to submission to bottom-up MS analysis. In this work, we repurpose a hydrogen-deuterium exchange MS (HDX-MS) LEAP HDX system for automated specific amino acid footprinting MS, demonstrating its efficacy in reaction optimization and monitoring applicability to specific ligand binding systems. We screened reagent conditions for two model ligand-binding systems and demonstrate the method's efficacy for measuring differences induced by ligand binding. Our p

SUBMITTER: Wagner ND 

PROVIDER: S-EPMC10482561 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature