title: Comparative Analysis of Archaeal Lipid-linked Oligosaccharides That Serve as Oligosaccharide Donors for Asn Glycosylation.
abstract: The glycosylation of asparagine residues is the predominant protein modification in all three domains of life. An oligosaccharide chain is preassembled on a lipid-phospho carrier and transferred onto asparagine residues by the action of a membrane-bound enzyme, oligosaccharyltransferase. The oligosaccharide donor for the oligosaccharyl transfer reaction is dolichol-diphosphate-oligosaccharide in Eukaryota and polyprenol-diphosphate-oligosaccharide in Eubacteria. The donor in some archaeal species was reportedly dolichol-monophosphate-oligosaccharide. Thus, the difference in the number of phosphate groups aroused interest in whether the use of the dolichol-monophosphate type donors is widespread in the domain Archaea. Currently, all of the archaeal species with identified oligosaccharide donors have belonged to the phylum Euryarchaeota. Here, we analyzed the donor structures of two species belonging to the phylum Crenarchaeota, Pyrobaculum calidifontis and Sulfolobus solfataricus, in addition to two species from the Euryarchaeota, Pyrococcus furiosus and Archaeoglobus fulgidus The electrospray ionization tandem mass spectrometry analyses confirmed that the two euryarchaeal oligosaccharide donors were the dolichol-monophosphate type and newly revealed that the two crenarchaeal oligosaccharide donors were the dolichol-diphosphate type. This novel finding is consistent with the hypothesis that the ancestor of Eukaryota is rooted within the TACK (Thaum-, Aig-, Cren-, and Korarchaeota) superphylum, which includes Crenarchaea. Our comprehensive study also revealed that one archaeal species could contain two distinct oligosaccharide donors for the oligosaccharyl transfer reaction. The A. fulgidus cells contained two oligosaccharide donors bearing oligosaccharide moieties with different backbone structures, and the S. solfataricus cells contained two oligosaccharide donors bearing stereochemically different dolichol chains.
authors.affiliation: From the Division of Structural Biology and.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Taguchi
authors.affiliation: From the Division of Structural Biology and.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Fujinami
authors.affiliation: From the Division of Structural Biology and Research Centers for Advanced Immunology and Live-Protein Dynamics, Medical Institute of Bioregulation, Kyushu University, Fukuoka 812-8582, Japan kohda@bioreg.kyushu-u.ac.jp.
authors.forename: Daisuke
authors.identifier: http://orcid.org/0000-0001-8234-3776
authors.initials: D
authors.lastname: Kohda
mesh_terms: D001105:Archaea / Q000145:classification / Q000378:metabolism*; D019843:Archaeal Proteins / Q000378:metabolism; D019611:Archaeoglobus fulgidus / Q000378:metabolism; D001216:Asparagine / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D006602:Hexosyltransferases / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D015394:Molecular Structure; D011104:Polyisoprenyl Phosphate Oligosaccharides / Q000737:chemistry* / Q000378:metabolism*; D041543:Pyrobaculum / Q000378:metabolism; D020027:Pyrococcus furiosus / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D048229:Sulfolobus solfataricus / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D019843:Archaeal Proteins; D008565:Membrane Proteins; D011104:Polyisoprenyl Phosphate Oligosaccharides; D001216:Asparagine; D006602:Hexosyltransferases; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase
keywords: AglB; N-linked glycosylation; archaea; crenarchaeota; dolichol; euryarchaeota; evolution; lipid-linked oligosaccharide; mass spectrometry (MS); oligosaccharyltransferase