The Neutral N ‐linked Glycans of the Ustilaginomycete Yeast Sympodiomycopsis paphiopedili

In this study, the neutral N‐linked glycans of S. paphiopedili were prepared and structurally analyzed using high performance liquid chromatography (HPLC) and mass spectrometry (MS). Glycosidase digestion analyses were also performed to verify certain glycan linkages. HPLC and MS analyses revealed the presence of neutral N‐linked glycans ranging from Man3GlcNAc2‐PA to Man9GlcNAc2‐PA in length. The most abundant neutral N‐linked glycan structure in this species was found to be the Manα1‐2Manα1‐6(Manα1‐3)Manα1‐6(Manα1‐2Manα1‐2Manα1‐3)Manβ1‐4GlcNAcβ1‐4GlcNAc (M8A). Moreover, the second and third most abundant neutral N‐linked glycan in S. paphiopedili were the Manα1‐2Manα1‐6(Manα1‐2Manα1‐3)Manα1‐6(Manα1‐2Manα1‐2Manα1‐3)Manβ1‐4GlcNAcβ1‐4GlcNAc (M9A) and the Manα1‐6(Manα1‐3)Manβ1‐4GlcNAcβ1‐4GlcNAc (M3B). On the other hand, the effect of the combination of glycoprotein extraction methods (citrate buffer extraction or beads extraction) and the subsequent glycan release methods (hydrazinolysis or PNGase F digestion) to the detection of N‐linked glycan peaks were also examined for S. paphiopedili and Saccharomyces cerevisiae in order to avoid under‐representation of N‐linked glycan structures. High mannose and possible hypermannosylated glycan peaks were detected in all method combinations in S. cerevisiae with the citrate buffer extraction‐hydrazinolysis method giving the highest peak yields as c...
Source: Yeast - Category: Molecular Biology Authors: Tags: Research Article Source Type: research