Project description:N-glycosylation is one of the most abundant and conserved protein modifications in eukaryotes. This modification serves various important functions, such as protein folding and cellular attachment, but also modulation of a protein’s function. Recently, it has been shown that N-glycosylation of proteins plays a vital role in insect development and survival, which makes it an interesting target for pest control. Despite the importance of protein N-glycosylation in insects, not much is known about insect N-glycoproteomes. Here, we report on the N-glycoproteomes of three major pest insects spanning different insect orders; Drosophila melanogaster (Diptera), Tribolium castaneum (Coleoptera) and Acyrthosiphon pisum (Hemiptera). The number of identified N-glycosylation sites ranged from 889 in T. castaneum, to 941 in D. melanogaster and 1,338 in A. pisum. Comparison between the different insect species revealed both conserved and species-specific glycoproteins. The functionality of the insect glycoproteins together with the conservation of the N-glycosites throughout evolution are discussed. This information can help in the elaboration of novel pest insect control strategies based on interference in insect glycosylation.