Probing Peptide Fragment Ion Structures by Combining Sustained Off-Resonance Collision-Induced Dissociation and Gas-Phase H/D Exchange (SORI-HDX) in Fourier Transform Ion-Cyclotron Resonance (FT-ICR) Instruments

The usefulness of gas-phase H/D exchange is demonstrated to probe heterogeneous fragment and parent ion populations. Singly and multiply protonated peptides/proteins were fragmented by using sustained off-resonance irradiation collision-induced dissociation (SORI-CID). The fragments and the surviving precursor ions then all undergo H/D exchange in the gas-phase with either D2O or CD3OD under the same experimental conditions. Usually, 10 to 60 s of reaction time is adequate to monitor characteristic differences in the H/D exchange kinetic rates. These differences are then correlated to isomeric ion structures. The SORI-HDX method can be used to rapidly test fragment ion structures and provides useful insights into peptide fragmentation mechanisms.

Graphical AbstractA gas-phase H/D exchange method is described and used to study structural differences in peptide fragments generated by SORI(RE)-CID in an FT-ICR instrument.Figure optionsDownload full-size imageDownload high-quality image (125 K)Download as PowerPoint slide