Mesh ： Amino Acid Sequence / genetics Amino Acid Substitution / genetics Amino Acids / chemistry genetics metabolism Bacterial Outer Membrane Proteins / chemistry genetics metabolism Bacterial Proteins / chemistry genetics metabolism Campylobacter jejuni / metabolism Consensus Sequence / genetics Glycoproteins / chemistry genetics metabolism Glycosylation Hexosyltransferases / metabolism Lipoproteins / chemistry genetics metabolism Membrane Proteins / metabolism Molecular Sequence Data Mutation Saccharomyces cerevisiae / metabolism

来  源：   DOI：10.1038/sj.emboj.7601087   PDF(Sci-hub)   PDF(Pubmed)

Abstract：
The Campylobacter jejuni pgl locus encodes an N-linked protein glycosylation machinery that can be functionally transferred into Escherichia coli. In this system, we analyzed the elements in the C. jejuni N-glycoprotein AcrA required for accepting an N-glycan. We found that the eukaryotic primary consensus sequence for N-glycosylation is N terminally extended to D/E-Y-N-X-S/T (Y, X not equalP) for recognition by the bacterial oligosaccharyltransferase (OST) PglB. However, not all consensus sequences were N-glycosylated when they were either artificially introduced or when they were present in non-C. jejuni proteins. We were able to produce recombinant glycoproteins with engineered N-glycosylation sites and confirmed the requirement for a negatively charged side chain at position -2 in C. jejuni N-glycoproteins. N-glycosylation of AcrA by the eukaryotic OST in Saccharomyces cerevisiae occurred independent of the acidic residue at the -2 position. Thus, bacterial N-glycosylation site selection is more specific than the eukaryotic equivalent with respect to the polypeptide acceptor sequence.
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