Definition of the bacterial N-glycosylation site consensus sequence

EMBO J. 2006 May 3;25(9):1957-66. doi: 10.1038/sj.emboj.7601087. Epub 2006 Apr 13.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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


  • Amino Acids
  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Glycoproteins
  • Lipoproteins
  • Membrane Proteins
  • Hexosyltransferases
  • dolichyl-diphosphooligosaccharide - protein glycotransferase