New N-glycans in horseradish peroxidase

Abstract

We describe here the structures of several new N-glycan oligosaccharides from horseradish peroxidase (HRP). Glycopeptides from HRP were digested with glycoamidase A (from sweet almond) to release the N-glycans. The oligosaccharides were reductively aminated with 2-aminopyridine, and separated by high-performance liquid chromatography on octadecylsilyl- and amide-silica columns for two-dimensional mapping (Tomiya et al., Anal. Biochem. 171, 73-90, 1988). The N-glycans were also analyzed by electrospray ionization mass spectrometry. Two different lots of HRP showed a considerable difference in their N-glycan composition.