N-glycoproteomic profiling of the hookworm Ancylostoma ceylanicum reveals heterogeneity and phosphorylcholine-modified glycans in larval and adult parasites.

Phosphorylcholine-modified N-glycans are detected in both larval and adult Ancylostoma ceylanicum, with stage-specific N-glycoproteins and unique PC-modified structures in each stage.

• Why it matters: Helminth glycans can modulate host immunity, but Ancylostoma ceylanicum glycosylation patterns are poorly defined due to sample-collection challenges and complex nematode glycan structures.
• What they did: Adult released N-glycan analysis identified oligomannose, hybrid, and complex structures, including non-mammalian features; glycoproteomics compared three enrichment methods, with SAX-ERLIC yielding the most N-glycopeptides.
• The result: Larval and adult extracts contained 82 larval and 157 adult N-glycoproteins, mostly stage-specific, and PC-modified glycans were found in both stages, enabling first site-specific characterization of intact N-glycopeptides in A. ceylanicum.