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

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 or on Thermo Fisher Orbitrap mass spectrometer Eclipse with ETD using HCD 204 triggered EtHCD.