Nakagawa S, Yagi H, Suyama T, Shimamura S, Yanaka S, Yagi-Utsumi M, Kato S, Ohkuma M, Kato K, Takai K

mBio, 16 (2025)

Ammonia-oxidizing archaea of the phylum Nitrososphaerota, formerly known as Thaumarchaeota, are globally distributed and play critical roles in the nitrogen and carbon cycles, particularly in environments with low ammonia concentrations. Like most archaea, Nitrososphaerota cells are enveloped by S-layer proteins, implicated in concentrating ammonium ions. These proteins are typically modified post-translationally by N-glycans, which often play significant roles in various biological processes, including protein function regulation, protection from phages, and environmental adaptation. Nevertheless, the glycobiological characteristics of Nitrososphaerota remain largely unexplored. Here, we investigated the glycoproteome of ammonia-oxidizing Nitrososphaerota, specifically focusing on the terrestrial Nitrososphaera viennensis and the marine Nitrosopumilus piranensis. Both species exhibited similar protein arrays throughout their growth phases, including those associated with N-glycosylation. Ns. viennensis consistently exhibited N-glycosylation predominantly on an S-layer protein and multicopper oxidase domain-containing proteins throughout all growth phases, with a marked increase during and after the late exponential phase. The glycan, characterized as a novel hexasaccharide with a chitobiose core, is hypothesized to play a role in nitrogen storage due to its probable nitrogen-rich composition, modifying asparagine residues within the conserved triplet sequence (Asn-X-Ser or -Thr). In contrast, Np. piranensis also showed a high abundance of S-layer protein but displayed no apparent N-glycosylation on any protein, suggesting variability in cell surface physical properties between these archaea. Despite similarities in their proteomes and energy metabolism, these two archaea exhibited significant differences in post-translational modification of proteins, revealing previously unrecognized diversity that may have implications for understanding their adaptive transitions to diverse environments.