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N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus.

Chang MM, Imperiali B, Eichler J, Guan Z - PLoS ONE (2015)

Bottom Line: In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized.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.

View Article: PubMed Central - PubMed

Affiliation: Departments of Biology and Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States of America.

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.

No MeSH data available.


The glycan-charged DolP species detected in the P. furiosus lipid extract.The predicted chemical structures and calculated masses of C65 DolP charged with the mono-, di-, tri-, tetra-, penta-, hexasaccharide-, and the complete heptasaccharide are shown (top to bottom, respectively).
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pone.0130482.g002: The glycan-charged DolP species detected in the P. furiosus lipid extract.The predicted chemical structures and calculated masses of C65 DolP charged with the mono-, di-, tri-, tetra-, penta-, hexasaccharide-, and the complete heptasaccharide are shown (top to bottom, respectively).

Mentions: In addition to heptasaccharide-charged DolP, as well as the hexa- and pentasaccharide-charged precursors, RPLC-ESI MS also revealed the presence of tetra-, tri-, di- and monosaccharide-charged DolP in the P. furiosus lipid extract. The chemical structures and exact masses of different glycan-charged C65 DolP species are presented in Fig 2.


N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus.

Chang MM, Imperiali B, Eichler J, Guan Z - PLoS ONE (2015)

The glycan-charged DolP species detected in the P. furiosus lipid extract.The predicted chemical structures and calculated masses of C65 DolP charged with the mono-, di-, tri-, tetra-, penta-, hexasaccharide-, and the complete heptasaccharide are shown (top to bottom, respectively).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476749&req=5

pone.0130482.g002: The glycan-charged DolP species detected in the P. furiosus lipid extract.The predicted chemical structures and calculated masses of C65 DolP charged with the mono-, di-, tri-, tetra-, penta-, hexasaccharide-, and the complete heptasaccharide are shown (top to bottom, respectively).
Mentions: In addition to heptasaccharide-charged DolP, as well as the hexa- and pentasaccharide-charged precursors, RPLC-ESI MS also revealed the presence of tetra-, tri-, di- and monosaccharide-charged DolP in the P. furiosus lipid extract. The chemical structures and exact masses of different glycan-charged C65 DolP species are presented in Fig 2.

Bottom Line: In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized.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.

View Article: PubMed Central - PubMed

Affiliation: Departments of Biology and Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States of America.

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.

No MeSH data available.