Functional characterization of bacterial oligosaccharyltransferases involved in O-linked protein glycosylation.

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, Neisseria gonorrhoeae, 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 oligosaccharide 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 lipid carriers to hydroxylated amino acids in proteins. To date, such activity has not been identified for eukaryotes. To our knowledge, this is the first report describing recombinant O glycoproteins synthesized in E. coli.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Faridmoayer A,Fentabil MA,Mills DC,Klassen JS,Feldman MF

doi

10.1128/JB.01318-07

subject

Has Abstract

pub_date

2007-11-01 00:00:00

pages

8088-98

issue

22

eissn

0021-9193

issn

1098-5530

pii

JB.01318-07

journal_volume

189

pub_type

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