Global-scale enrichment and identification of O-GlcNAc modified peptides in Arabidopsis seedling (S1 Part 1: HCD and EThcD part from Eclipse)

O-GlcNAcylation is a crucial post-translational modification of proteins observed in both plants and animals and plays a key role in growth and development. However, our understanding of this PTM in plants lags behind animals. To bridge this knowledge gap, we performed a global scale enrichment of O-GlcNAc modified peptides after LWAC enrichment on Col seedlings. A 15N-labeled spy-4 is also included in the samples for comparison. Enriched samples were subjected to extensive reverse high pH fractionation followed by LC-MS-MS analysis on Thermo Fisher Orbitrap mass spectrometer (QExactive HF) using HCD modes (Files initiate with Q) or on Thermo Fisher Orbitrap mass spectrometer Eclipse with ETD using HCD 204 triggered EtHCD.