Surface-exposed Glycoproteins of Hyperthermophilic Sulfolobus solfataricus P2 Show a Common N-Glycosylation Profile
datasetposted on 07.06.2013, 00:00 authored by Gianna Palmieri, Marco Balestrieri, Jasna Peter-Katalinić, Gottfried Pohlentz, Mosè Rossi, Immacolata Fiume, Gabriella Pocsfalvi
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.
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sulfated heptasaccharidesurface fractionintercellular communicationglycoproteinlectin affinity purificationenergy conversionarchaeal glycan structurehyperthermophilic Archaeasurvival strategiesglycan compositionsSSO 1273 proteinnovel glycan structuremonosaccharide unitsulfated oligosaccharideHyperthermophilic Sulfolobus solfataricus P 2 Showconsensus sequence sitescell surface fractionSulfolobus solfataricus P 2archaeal speciesABC transporters