Fast Protein Footprinting by X-ray Mediated Radical Trifluoromethylation

Synchrotron radiolysis generates hydroxyl radicals (^•OH) that are successful footprinting reagents. Here, we describe a new reagent for the synchrotron platform, the trifluoromethyl radical (^•CF₃). The radical is produced by ^•OH displacement of ^•CF₃ from sodium triflinate (Langlois reagent). Upon X-ray beam exposure, the reagent labels proteins extensively without any additional chemicals on a millisecond or shorter time scale. The ^•CF₃ is comparably reactive to ^•OH and produces footprinting information that complements that of ^•OH alone. This reagent in combination with ^•OH should enable novel chemistry for protein footprinting on the synchrotron platform.