title: N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus.
abstract: In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated.
authors.affiliation: Departments of Biology and Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States of America.
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Chang
authors.affiliation: Departments of Biology and Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States of America.
authors.forename: Barbara
authors.initials: B
authors.lastname: Imperiali
authors.affiliation: Department of Life Sciences, Ben Gurion University, Beersheva 84105, Israel.
authors.forename: Jerry
authors.initials: J
authors.lastname: Eichler
authors.affiliation: Department of Biochemistry, Duke University Medical Center, Durham NC 27710, United States of America.
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Guan
mesh_terms: D001105:Archaea / Q000378:metabolism*; D002070:Butadienes / Q000378:metabolism; D004288:Dolichol Phosphates / Q000378:metabolism*; D004286:Dolichols / Q000378:metabolism*; D006031:Glycosylation; D045782:Hemiterpenes / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010420:Pentanes / Q000378:metabolism; D028061:Phosphate Transport Proteins / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D020027:Pyrococcus furiosus / Q000378:metabolism*
chemical_list: D002070:Butadienes; D004288:Dolichol Phosphates; D004286:Dolichols; D045782:Hemiterpenes; D009844:Oligosaccharides; D010420:Pentanes; D028061:Phosphate Transport Proteins; D011134:Polysaccharides; C005059:isoprene; C028306:dolichol monophosphate; C026406:dolichol pyrophosphate