Abstract
N-glycosylation is one of the most abundant and conserved protein modifications in eukaryotes. The attachment of N-glycans to proteins can modulate their properties and influences numerous important biological processes, such as protein folding and cellular attachment. Recently, it has been shown that protein N-glycosylation plays a vital role in insect development and survival, which makes the glycans an interesting target for pest control. Despite the importance of protein N-glycosylation in insects, knowledge about insect N-glycoproteomes is scarce. To fill this gap, the N-glycosites were identified in proteins from three major pest insects, spanning different insect orders and diverging in post-embryonic development, feeding mechanism and evolutionary ancestry: Drosophila melanogaster (Diptera), Tribolium castaneum (Coleoptera) and Acyrthosiphon pisum (Hemiptera). The N-glyco-FASP method for isolation of N-glycopeptides was optimized to study the insect N-glycosites and allowed the identification of 889 N-glycosylation sites in T. castaneum, 941 in D. melanogaster and 1338 in A. pisum. Although a large set of the corresponding glycoproteins is shared among the three insects, species- and order-specific glycoproteins were also identified. The functionality of the insect glycoproteins together with the conservation of the N-glycosites throughout evolution is discussed. This information can help in the elaboration of novel pest insect control strategies based on interference in insect glycosylation.
