Multiplex Trifluoromethyl and Hydroxyl Radical Chemistry Enables High-Resolution Protein Footprinting.

ABSTRACT: Hydroxyl radical-based protein footprinting (HRPF) coupled with mass spectrometry is a valuable medium-resolution technique in structural biology, facilitating the assessment of protein structure and molecular-level interactions in solution conditions. In HRPF with X-rays (XFP), hydroxyl radicals generated by water radiolysis covalently label multiple amino acid (AA) side chains. However, HRPF technologies face challenges in achieving their full potential due to the broad (>10³) dynamic range of AA reactivity with ^•OH and difficulty in detecting slightly modified residues, most notably in peptides with highly reactive residues like methionine, or where all residues have low ^•OH reactivities. To overcome this limitation, we developed a multiplex labeling che

SUBMITTER: Jain R 

PROVIDER: S-EPMC11830425 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature