This Article Full Text (PDF) Other Versions of this Article: JB.01318-07v1 189/22/8088    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by FARIDMOAYER, A. Articles by FELDMAN, M. F. Search for Related Content PubMed PubMed Citation Articles by FARIDMOAYER, A. Articles by FELDMAN, M. F.

 Previous Article  |  Next Article 

J. Bacteriol. doi:10.1128/JB.01318-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Functional Characterization of Bacterial Oligosaccharyltransferases Involved in O-linked Protein Glycosylation

Alberta Ingenuity Centre for Carbohydrate Science, Department of Biological Sciences, Department of Chemistry, University of Alberta, T6G 2E9, Edmonton, Alberta, Canada

^* To whom correspondence should be addressed. Email: mfeldman{at}ualberta.ca.

	Abstract

Protein glycosylation is an important posttranslational modification that occurs in all domains of life. Pilins, the structural components of type IV pili, are O-glycosylated in Neisseria meningitidis, N. gonorrhea, and some strains of Pseudomonas aeruginosa. In this work, we characterized the P. aeruginosa 1244 and N. meningitidis MC58 O-glycosylation systems in Escherichia coli. In both cases sugars are transferred en bloc by an oligosaccharyltransferase (OTase), named PglL in N. meningitidis and PilO in P. aeruginosa. We show that, like PilO, PglL has relaxed glycan specificity. Both OTases are sufficient for glycosylation but they require translocation of the undecaprenol-pyrophosphate–linked oligosaccharides substrates into the periplasm for activity. Whereas PilO activity is restricted to short oligosaccharides, PglL is able to transfer diverse oligo- and polysaccharides. This functional characterization supports the concept that despite their low sequence similarity, PilO and PglL belong to a new family of "O-OTases" that transfer oligosaccharides from a lipid carrier to hydroxylated amino acids in proteins. To date, such activity has not been identified in eukaryotes. To our knowledge, this is the first report describing recombinant O-glycoproteins synthesized in E. coli.

This article has been cited by other articles:

• Steiner, K., Novotny, R., Werz, D. B., Zarschler, K., Seeberger, P. H., Hofinger, A., Kosma, P., Schaffer, C., Messner, P. (2008). Molecular Basis of S-layer Glycoprotein Glycan Biosynthesis in Geobacillus stearothermophilus. J. Biol. Chem. 283: 21120-21133 [Abstract] [Full Text]