Oral Presentation 31st Annual Lorne Proteomics Symposium 2026

The diversity and evolution of protein N-glycosylation in Metazoa (133408)

Fabian Ruperti 1 , Clement M Potel 2 , Isabelle Becher 2 , Leslie Pan 2 , Sandie Degnan 1 , Bernard Degnan 1 , Detlev Arendt 2 , Ben Schulz 1 , Mikhail M Savitski 2
  1. University of Queensland, Brisbane, St. Lucia, QUEENSLAND, Australia
  2. European Molecular Biology Laboratory, Heidelberg, Germany

N-linked glycosylation is a complex post-translational modification in which (oligo)saccharides are covalently attached to extracellular and membrane-bound proteins. As such, N-glycoproteins are central to cell-cell and cell-ECM adhesion, recognition and communication, making them fundamental players in key animal traits such as multicellularity, the immune and nervous systems, and interactions with symbionts [1]. Despite their importance, our understanding of the diversity, conservation, heterogeneity and role of N-glycoproteins and their attached glycans in animal evolution is currently poor, owing to sparse taxonomic sampling and the lack of universal analytical frameworks.

Here, we apply high-throughput, high-resolution mass spectrometry-based glycoproteomics [2] to profile the N-glycoproteomes of ~30 opisthokont species spanning major evolutionary transitions from unicellular fungi and holozoans to non-bilaterian and bilaterian animals. To resolve glycan class diversity across distant lineages, we developed an empirical, FDR-controlled, diagnostic ion-informed annotation pipeline that operates on open-search results for each individual species. This enables robust glycan class assignment, while providing the capacity to discover, curate, and validate cryptic diagnostic ions. In doing so, we provide the first site- and glycan-resolved N-glycoproteome resource for multiple phyla, including choanoflagellates, ctenophores, sponges, and cnidarians, while also greatly expanding coverage for well-studied model species. Our results show that the biochemical logic of N-glycosylation sites is deeply conserved, yet the repertoire of attached glycans is remarkably diverse, showing lineage-specific innovations and convergence across distantly related clades. Ongoing comparative analyses across orthologous glycoproteins aim to trace the emergence and conservation of glycosylation sites, glycan classes and their heterogeneity, and link these features to protein function and cell type-specificity.

Together, the analysis of this expansive new resource will reveal how changes in the “glycan code of life” contributed to the origin of animal multicellularity, the diversification of cell types and their intra- and interspecies interactions.



  1. [1] Tivey, Trevor R., et al. "N-linked surface glycan biosynthesis, composition, inhibition, and function in cnidarian-dinoflagellate symbiosis." Microbial ecology 80.1 (2020): 223-236.
  2. [2] Potel, Clément M., et al. "Uncovering protein glycosylation dynamics and heterogeneity using deep quantitative glycoprofiling (DQGlyco)." Nature Structural & Molecular Biology 32.6 (2025): 1111-1126.