Dolichol-linked oligosaccharide selection by the oligosaccharyltransferase in protist and fungal organisms

Daniel J. Kelleher, Sulagna Banerjee, Anthony J. Cura, John Samuelson, Reid Gilmore

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

The dolichol-linked oligosaccharide Glc3Man9Glc- NAc2-PP-Dol is the in vivo donor substrate synthesized by most eukaryotes for asparagine-linked glycosylation. However, many protist organisms assemble dolichol-linked oligosaccharides that lack glucose residues. We have compared donor substrate utilization by the oligosaccharyltransferase (OST) from Trypanosoma cruzi, Entamoeba histolytica, Trichomonas vaginalis, Cryptococcus neoformans, and Saccharomyces cerevisiae using structurally homogeneous dolichol-linked oligosaccharides as well as a heterogeneous dolichol-linked oligosaccharide library. Our results demonstrate that the OST from diverse organisms utilizes the in vivo oligo saccharide donor in preference to certain larger and/or smaller oligosaccharide donors. Steady-state enzyme kinetic experiments reveal that the binding affinity of the tripeptide acceptor for the protist OST complex is influenced by the structure of the oligosaccharide donor. This rudimentary donor substrate selection mechanism has been refined in fungi and vertebrate organisms by the addition of a second, regulatory dolichol-linked oligosaccharide binding site, the presence of which correlates with acquisition of the SWP1/ribophorin II subunit of the OST complex.

Original languageEnglish (US)
Pages (from-to)29-37
Number of pages9
JournalJournal of Cell Biology
Volume177
Issue number1
DOIs
StatePublished - Apr 9 2007
Externally publishedYes

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Dolichol
Oligosaccharides
Donor Selection
Trichomonas vaginalis
Entamoeba histolytica
Cryptococcus neoformans
Trypanosoma cruzi
Asparagine
dolichyl-diphosphooligosaccharide - protein glycotransferase
Eukaryota
Glycosylation
Saccharomyces cerevisiae
Vertebrates
Fungi
Binding Sites
Glucose
Enzymes

ASJC Scopus subject areas

  • Cell Biology

Cite this

Dolichol-linked oligosaccharide selection by the oligosaccharyltransferase in protist and fungal organisms. / Kelleher, Daniel J.; Banerjee, Sulagna; Cura, Anthony J.; Samuelson, John; Gilmore, Reid.

In: Journal of Cell Biology, Vol. 177, No. 1, 09.04.2007, p. 29-37.

Research output: Contribution to journalArticle

Kelleher, Daniel J. ; Banerjee, Sulagna ; Cura, Anthony J. ; Samuelson, John ; Gilmore, Reid. / Dolichol-linked oligosaccharide selection by the oligosaccharyltransferase in protist and fungal organisms. In: Journal of Cell Biology. 2007 ; Vol. 177, No. 1. pp. 29-37.
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