Department of Biochemistry and Molecular Pharmacology
Animals; Binding Sites; Cryptococcus neoformans; Dolichol; Entamoeba histolytica; Fungal Proteins; Hexosyltransferases; Kinetics; Mannose; Membrane Proteins; Oligosaccharides; Protozoan Proteins; Saccharomyces cerevisiae; Substrate Specificity; Trichomonas vaginalis; Trypanosoma cruzi
Life Sciences | Medicine and Health Sciences
The dolichol-linked oligosaccharide Glc3Man9GlcNAc2-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.
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Citation: J Cell Biol. 2007 Apr 9;177(1):29-37. Epub 2007 Apr 2. Link to article on publisher's site
DOI of Published Version
The Journal of cell biology glycotransferase)
Kelleher, Daniel J.; Banerjee, Sulagna; Cura, Anthony J.; Samuelson, John C.; and Gilmore, Reid, "Dolichol-linked oligosaccharide selection by the oligosaccharyltransferase in protist and fungal organisms" (2007). Open Access Articles. 1280.