Mass spectrometry detects folding intermediates populated during urea-induced protein denaturation.

ABSTRACT: Protein folding stability can be probed using urea, a chaotropic agent that disrupts non-covalent interactions at molar concentrations. The denaturation process is typically monitored via optical spectroscopy, which provides ensemble-averaged measurements and may struggle to resolve folding intermediates. In contrast, electrospray ionization mass spectrometry (ESI-MS) captures a non-averaged snapshot of all populated assembly and folding states within a protein conformational ensemble. However, high urea concentrations have traditionally been considered incompatible with ESI. Here, we leverage recent advancements in nano ESI emitter design, utilizing well-defined small-diameter emitters which enables protein charge states to be resolved from solutions containing up to 8 M urea. This

SUBMITTER: Osterlund N 

PROVIDER: S-EPMC12573884 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature