Q-TOF LC/MS analysis of GspB736flag after trypsin, Lys-C, and Glu-C protease digestion.
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Extracted ion chromatograms for all detected glycopeptides derived from GspB736flag secreted from A) a Δgly-nss strain (PS3309) or B) a Δgly-nss Asp2S362A strain (PS3540). Peaks with the same peptide backbone are designated with the same color (green, red, black and blue, respectively) and asterisks denote those containing an O-acetylated glycopeptide. C) Fragmentation spectrum of peak 1, m/z = 484.73 [M+2H]2+ O-glycosylated at two positions. D) Fragmentation spectrum of peak 1*, m/z = 505.74 [M+2H]2+ O-glycosylated at two positions and O-acetylated at one position. O-GlcNAc modifications are designated by a blue square. MS spectra of precursor ions are shown in the insets.