Comprehensive glycoproteomics shines new light on the complexity and extent of glycosylation in archaea

PLoS Biol. 2021 Jun 17;19(6):e3001277. doi: 10.1371/journal.pbio.3001277. eCollection 2021 Jun.

Abstract

Glycosylation is one of the most complex posttranslational protein modifications. Its importance has been established not only for eukaryotes but also for a variety of prokaryotic cellular processes, such as biofilm formation, motility, and mating. However, comprehensive glycoproteomic analyses are largely missing in prokaryotes. Here, we extend the phenotypic characterization of N-glycosylation pathway mutants in Haloferax volcanii and provide a detailed glycoproteome for this model archaeon through the mass spectrometric analysis of intact glycopeptides. Using in-depth glycoproteomic datasets generated for the wild-type (WT) and mutant strains as well as a reanalysis of datasets within the Archaeal Proteome Project (ArcPP), we identify the largest archaeal glycoproteome described so far. We further show that different N-glycosylation pathways can modify the same glycosites under the same culture conditions. The extent and complexity of the Hfx. volcanii N-glycoproteome revealed here provide new insights into the roles of N-glycosylation in archaeal cell biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Archaeal Proteins / chemistry
  • Archaeal Proteins / metabolism*
  • Biological Assay
  • Cell Shape / drug effects
  • Databases, Protein
  • Glycopeptides / chemistry
  • Glycopeptides / metabolism*
  • Glycoproteins / chemistry
  • Glycoproteins / metabolism*
  • Glycosylation / drug effects
  • Haloferax volcanii / drug effects
  • Haloferax volcanii / metabolism*
  • Mutation / genetics
  • Phenotype
  • Phylogeny
  • Proteomics
  • Sodium Chloride / pharmacology

Substances

  • Archaeal Proteins
  • Glycopeptides
  • Glycoproteins
  • Sodium Chloride