Surface-exposed glycoproteins of hyperthermophilic Sulfolobus solfataricus P2 show a common N-glycosylation profile

J Proteome Res. 2013 Jun 7;12(6):2779-90. doi: 10.1021/pr400123z. Epub 2013 Apr 30.

Abstract

Cell surface proteins of hyperthermophilic Archaea actively participate in intercellular communication, cellular uptake, and energy conversion to sustain survival strategies in extreme habitats. Surface (S)-layer glycoproteins, the major component of the S-layers in many archaeal species and the best-characterized prokaryotic glycoproteins, were shown to have a large structural diversity in their glycan compositions. In spite of this, knowledge on glycosylation of proteins other than S-layer proteins in Archaea is quite limited. Here, the N-glycosylation pattern of cell-surface-exposed proteins of Sulfolobus solfataricus P2 were analyzed by lectin affinity purification, HPAEC-PAD, and multiple mass spectrometry-based techniques. Detailed analysis of SSO1273, one of the most abundant ABC transporters present in the cell surface fraction of S. solfataricus, revealed a novel glycan structure composed of a branched sulfated heptasaccharide, Hex4(GlcNAc)2 plus sulfoquinovose where Hex is d-mannose and d-glucose. Having one monosaccharide unit more than the glycan of the S-layer glycoprotein of S. acidocaldarius, this is the most complex archaeal glycan structure known today. SSO1273 protein is heavily glycosylated and all 20 theoretical N-X-S/T (where X is any amino acid except proline) consensus sequence sites were confirmed. Remarkably, we show that several other proteins in the surface fraction of S. solfataricus are N-glycosylated by the same sulfated oligosaccharide and we identified 56 N-glycosylation sites in this subproteome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / chemistry
  • ATP-Binding Cassette Transporters / isolation & purification*
  • Amino Acid Sequence
  • Archaeal Proteins / chemistry
  • Archaeal Proteins / isolation & purification*
  • Carbohydrate Sequence
  • Chromatography, Affinity
  • Glycosylation
  • Lectins / chemistry
  • Mass Spectrometry
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / isolation & purification*
  • Molecular Sequence Data
  • Oligosaccharides, Branched-Chain / chemistry
  • Oligosaccharides, Branched-Chain / isolation & purification*
  • Sulfolobus solfataricus / chemistry*

Substances

  • ATP-Binding Cassette Transporters
  • Archaeal Proteins
  • Lectins
  • Membrane Glycoproteins
  • Oligosaccharides, Branched-Chain