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Definition of the bacterial N‐ glycosylation site consensus sequence
Author(s) -
Kowarik Michael,
Young N Martin,
Numao Shin,
Schulz Benjamin L,
Hug Isabelle,
Callewaert Nico,
Mills Dominic C,
Watson David C,
Hernandez Marcela,
Kelly John F,
Wacker Michael,
Aebi Markus
Publication year - 2006
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1038/sj.emboj.7601087
Subject(s) - biology , consensus sequence , glycosylation , sequence (biology) , computational biology , bacterial protein , genetics , peptide sequence , sequence alignment , bacteria , gene
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≠P) 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.