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. 2013 Apr 12;32(10):1478–1488. doi: 10.1038/emboj.2013.79

Figure 1.

Figure 1

The SimpleCell O-GalNAc glycoproteomics strategy. (A) SimpleCell lines originating from different organs as illustrated were generated by ZFN-mediated knockout of COSMC. (B) SC express homogeneous truncated O-glycans (Tn/STn), and glycopeptides from cell lysates as well as conditioned media can be isolated by LWAC after protease digestion with trypsin or chymotrypsin. If necessary, sialic acids are removed by neuraminidase treatment. GalNAc-glycopeptides are isolated on LWAC and separated by IEF prior to nLC-MS/MS analysis. *Glycoproteins from conditioned media were pre-concentrated by passing the media over a short VVA column (see Extended Experimental Procedures). (C) The SimpleCell strategy identified 629 glycoproteins (not including possible GlcNAc cases), with only 48 in common with glycoproteins previously reported in UniProt (106 total) and O-GLYCBASE (78 total). Of the 771 glycoproteins in these three data sets, only 526 are predicted to have a signal peptide by the SignalP predictor.