Review Volume 1, Issue 1 pp 17—27

Figure 1. Conservation of N-glycosylation sites in apoD among species. (A). ApoD protein sequences of different species were aligned using the ClustalW program. Amino acid residues in box denote two highly conserved N-glycosylation sites in apoD. The consensus N-glycosylation site is Asn-X-Ser/Thr. (B) Plasma apoD is N-glycosylated. Aliquots of plasma (20 μg protein) from C57BL/6J mice were incubated without (-) and with (+) 1,000 U of N-glycosidase F (New England Biolabs) in a total volume of 30 μl at 37°C for 1 hour to remove N-glycan chains from glycopeptides. The reaction mixture was resolved on 4-20% SDS-polyacrylamide gels, followed by immunoblot analysis using anti-apoD. Glycosylated and de-glycosylated forms of apoD are indicated.