Enantioselective Protein Affinity Selection Mass Spectrometry (E-ASMS).

ABSTRACT: We report an enantioselective protein affinity selection mass spectrometry screening approach (E-ASMS) that enables the detection of weak binders, informs on selectivity, and generates orthogonal confirmation of binding. After method development with control proteins, we screened 31 human proteins against a designed library of 8,210 chiral compounds. 16 binders to 12 targets, including many proteins predicted to be "challenging to ligand", were discovered and confirmed in orthogonal biophysical assays. 7 binders to 6 targets bound in an enantioselective manner, with K _D values ranging from 3 to 20 μM. Binders for four targets (DDB1, WDR91, WDR55, and HAT1) were selected for in-depth characterization using X-ray crystallography. In all four cases, the mechanism for enantio

SUBMITTER: Wang X 

PROVIDER: S-EPMC11785093 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature