Lewis X biosynthesis in Helicobacter pylori. Molecular cloning of an alpha(1,3)-fucosyltransferase gene.
The lipopolysaccharide of certain strains of Helicobacter pylori was recently shown to contain the Lewis X (Lex) trisaccharide (Galbeta-1, 4-(Fucalpha(1,3))-GlcNAc). Lex is an oncofetal antigen which appears on human gastric epithelium, and its mimicry by carbohydrate structures on the surface of H. pylori may play an important part in the interaction of this pathogen with its host. Potential roles for bacterial Lex in mucosal adhesion, immune evasion, and autoantibody induction have been proposed (Moran, A. P., Prendergast, M. M., and Appelmelk, B. J. (1996) FEMS Immunol. Med. Microbiol. 16, 105-115). In mammals, the final step of Lex biosynthesis is the alpha(1,3)-fucosylation of GlcNAc in a terminal Galbeta(1-->4)GlcNAc unit, and a corresponding GDP-fucose:N-acetylglucosaminyl alpha(1,3) fucosyltransferase (alpha(1,3)-Fuc-T) activity was recently discovered in H. pylori extracts. We used part of a human alpha(1, 3)-Fuc-T amino acid sequence to search an H. pylori genomic data base for related sequences. Using a probe based upon weakly matching data base sequences, we retrieved clones from a plasmid library of H. pylori DNA. DNA sequence analysis of the library clones revealed a gene which we have named fucT, encoding a protein with localized homology to the human alpha(1,3)-Fuc-Ts. We have demonstrated that fucT encodes an active Fuc-T enzyme by expressing the gene in Escherichia coli. The recombinant enzyme shows a strong preference for type 2 (e.g. LacNAc) over type 1 (e.g. lacto-N-biose) acceptors in vitro. Certain residues in a short segment of the H. pylori protein are completely conserved throughout the alpha(1,3)-Fuc-T family, defining an alpha(1,3)-Fuc-T motif which may be of use in identifying new fucosyltransferase genes.
Martin
S L
SL
Glycobiology Unit, GlaxoWellcome Medicines Research Centre, Stevenage, Herts SG1 2NY, United Kingdom. slm17885@ggr.co.uk
Edbrooke
M R
MR
Hodgman
T C
TC
van den Eijnden
D H
DH
Bird
M I
MI
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics*; D016256:Lewis X Antigen / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D004269:DNA, Bacterial; D016256:Lewis X Antigen; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase