Methionine Oxidation Footprinting in Intact Proteins (MOFIP) Using Top-Down Proteomics.

ABSTRACT: Mass spectrometry (MS)-based proteomics methods, including protein footprinting methods such as hydrogen-deuterium exchange mass spectrometry (HDX-MS) and hydroxyl radical footprinting (HRF), can give unique insight into protein structure and interactions. These methods primarily utilize bottom-up proteomics techniques that require the digestion of intact proteins into small peptides before MS analysis. This digestion can obscure structural information relevant to the function of the intact proteoforms. Here, we have developed a novel top-down footprinting method, Methionine Oxidation Footprinting in Intact Proteins (MOFIP), to probe solvent accessibility in intact proteoforms. For MOFIP, natively folded protein lysates are incubated with and without hydrogen peroxide (H₂O₂

SUBMITTER: Sunny AT 

PROVIDER: S-EPMC12788183 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature