Structural elucidation of an asparagine-linked oligosaccharide from the hyperthermophilic archaeon, Pyrococcus furiosus

An oligosaccharide chain attached to the asparagine residue in a structurally defined peptide was produced by an in vitro oligosaccharide-transfer reaction, using membrane fractions that contained the oligosaccharyltransferase from the hyperthermophilic archaeon, Pyrococcus furiosus. The chemical structure of the N-glycan was elucidated by sugar analysis, NMR spectroscopy, and MS spectrometry, which revealed the structure. alpha-D-ManpNAc-(1 -> 4)-alpha-D-GlcAp-(1 -> 3)-alpha-D-Manp-(1 -> 2)-alpha-D-Manp-(1 -> 3)-beta-D-GalpNAc-Asm 3 2 up arrow up arrow 1 1 beta-D-Xylp beta-D-Xylp The shorter glycan structures lacking one or two xylose residues were also transferred by the P. furiosus oligosaccharyltransferase. The archaeal N-glycans are known to exhibit a high degree of structural variation. The structure of the P. furiosus N-glycan is novel and unique. The present data will be useful for structural and functional studies of the P. furiosus oligosaccharyltransferase. (C) 2014 Elsevier Ltd. All rights reserved.