JCB logo
Accuri Cytometers
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
Published online April 2, 2007
doi:10.1083/jcb.200611079
The Journal of Cell Biology, Vol. 177, No. 1, 29-37
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Kelleher et al.
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 1566K)
Right arrow PPT slides of all figures
Right arrow Supplemental Material Index
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kelleher, D. J.
Right arrow Articles by Gilmore, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kelleher, D. J.
Right arrow Articles by Gilmore, R.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Article

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



Daniel J. Kelleher1, Sulagna Banerjee2, Anthony J. Cura1, John Samuelson2, and Reid Gilmore1

1 Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605
2 Department of Molecular and Cell Biology, Goldman School of Dental Medicine, Boston University, Boston, MA 02118

Correspondence to Reid Gilmore: reid.gilmore{at}umassmed.edu

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.

Abbreviations used in this paper: ALG, asparagine-linked glycosylation; HPLC, high-pressure liquid chromatography; OS-NYT, glycosylated tripeptide; OS-PP-Dol; dolichol-linked oligosaccharide; OST, oligosaccharyltransferase; PIC, protease inhibitor cocktail.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents