Fig. 4: Multi-dimension quantitative N-glycoproteomics of 2FF-treated human Burkitt’s lymphoma cells.

a Schematic explanation of the propagation of quantitative information from GPSMs to unique glycoforms, glycosites, and glycans in this study. For one N-glycosylation site (N) on a protein (P), multiple glycans (G) may exist on the site, resulting in multiple glycoforms (represented as P-N-G). Quantification of unique glycoforms are achieved by summing TMT-reporter ion intensities of all involving GPSMs resulted from miscleavages, modifications, and different charge states. Quantified glycoforms are further combined for glycosite (P-N) quantification. As for a unique glycan, the quantification values of all glycoforms with this particular glycan in the sample are combined. b The SugarQuant workflow conducted for the 2FF treatment on DG75 cells. n = 2 biologically independent cells. c Aligned ratio distributions of glycoforms quantified via the Glyco-SPS-MS3 (left) or the MS2 method (right) upon 2FF treatment. Different 2FF concentrations were color coded as shown in the figure. For direct comparison, we separate fucosylated glycoforms (upper panel) from non-fucosylated ones (lower panel). d, e Similar to b, but showing the 2FF-induced changes at the levels of glycosites (left) or glycan composition (right) determined by the Glyco-SPS-MS3. Source data are provided as a Source Data file.