Helicobacter pylori 1,3-N-acetylglucosaminyltransferase for versatile synthesis of type 1 and type 2 poly-LacNAcs on N-linked, O-linked and I-antigen glycans

Download
  1. Get@NRC: Helicobacter pylori 1,3-N-acetylglucosaminyltransferase for versatile synthesis of type 1 and type 2 poly-LacNAcs on N-linked, O-linked and I-antigen glycans (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1093/glycob/cws101
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleGlycobiology
ISSN0959-6658
Volume22
Issue11
Pages14531464; # of pages: 12
Subjectbeta 3 n acetylglucosaminyltransferase; glycan derivative; glycopeptide; n acetylglucosamine; n acetylglucosaminyltransferase; unclassified drug; aminosugar; blood group I antigen; glycosphingolipid; I-antigen; n acetylglucosamine; n acetyllactosamine; N-acetyllactosamine; polylactosamine; polysaccharide; article; carbon nuclear magnetic resonance; enzyme activity; enzyme substrate complex; Helicobacter pylori; priority journal; protein motif; proton nuclear magnetic resonance; synthesis; biosynthesis; chemistry; enzymology; glycosylation; metabolism; Bacteria (microorganisms); Helicobacter pylori; Acetylglucosamine; Amino Sugars; Glycosphingolipids; Glycosylation; Helicobacter pylori; N-Acetylglucosaminyltransferases; Polysaccharides
AbstractPoly-N-acetyllactosamine extensions on N-and O-linked glycans are increasingly recognized as biologically important structural features, but access to these structures has not been widely available. Here, we report a detailed substrate specificity and catalytic efficiency of the bacterial β3-N-acetylglucosaminyltransferase (β3GlcNAcT) from Helicobacter pylori that can be adapted to the synthesis of a rich diversity of glycans with poly-LacNAc extensions. This glycosyltransferase has surprisingly broad acceptor specificity toward type-1,-2,-3 and-4 galactoside motifs on both linear and branched glycans, found commonly on N-linked, O-linked and I-antigen glycans. This finding enables the production of complex ligands for glycan-binding studies. Although the enzyme shows preferential activity for type 2 (Galβ1-4GlcNAc) acceptors, it is capable of transferring N-acetylglucosamine (GlcNAc) in β1-3 linkage to type-1 (Galβ1-3GlcNAc) or type-3/4 (Gal1-3GalNAc/) sequences. Thus, by alternating the use of the H. pylori β3GlcNAcT with galactosyltransferases that make the 1-4 or 1-3 linkages, various N-linked, O-linked and I-antigen acceptors could be elongated with type-2 and type-1 LacNAc repeats. Finally, one-pot incubation of di-LacNAc biantennary N-glycopeptide with the β3GlcNAcT and GalT-1 in the presence of uridine diphosphate (UDP)-GlcNAc and UDP-Gal, yielded products with 15 additional LacNAc units on the precursor, which was seen as a series of sequential ion peaks representing alternative additions of GlcNAc and Gal residues, on matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Overall, our data demonstrate a broader substrate specificity for the H. pylori β3GlcNAcT than previously recognized and demonstrate its ability as a potent resource for preparative chemo-enzymatic synthesis of complex glycans. © 2012 The Author.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Biological Sciences (IBS-ISB)
Peer reviewedYes
NPARC number21269383
Export citationExport as RIS
Report a correctionReport a correction
Record identifier029c1d00-b51d-4472-975f-d71c960a4c3a
Record created2013-12-12
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)