Secreted as well as membrane-associated eukaryotic proteins are most commonly glycosylated. Saccharides are attached to proteins mainly through N-and O-glycosydic bonds or as part of the glycosylphosphatidyinositol-membrane anchor. In contrast to N-glycosylation, which involves the co-translational transfer in the endoplasmic reticulum (ER) of the glycan portion of Glc3Man9GlcNAc2-PP-dolichol to suitable Asn residues on nascent polypeptides, O-glycosylation begins with the addition of a single monosaccharide. Contrary to N-glycosylation, which involves an asparagine residue in the sequon Asn-Xaa-Thr/Ser (Xaa can be any amino acid except Pro, and it is rarely Cys), no particular sequence motif has been described for O-glycosylation. This may reflect the fact that: (1) the specificity of the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase is presently unknown; and (2) seems to be modulated by sequence context, secondary structure, and surface accessibility ( 1 ). An internet server, accessible at http://www.cbs.dtu.dk/netOglyc/cbsnetOglyc.html, produces neural network predictions of mucin-type GalNAc O-glycosylation sites in mammalian proteins based on 299 known and verified mucin-type O-glycosylation sites. The sequence context of glycosylated threonines was found to differ from that of serine, and the sites were found to cluster. Nonclustered sites had a sequence context different from that of clustered sites, and charged residues were disfavored at position −1 and +3.