Helicobacter pylori lipopolysaccharides (LPS) contain complex carbohydrates known as Lewis antigens which may contribute to the pathogenesis and adaptation of the bacterium. Involved in the biosynthesis of Lewis antigens is an α1,2-fucosyltransferase (FucT) that adds fucose to the terminal βGal unit of the O-chain of LPS. Recently, the H. pylori (Hp) α1,2-FucT-encoding gene (fucT2) was cloned and analysed in detail. However, due to the low level of expression and instability of the protein, its enzymic activity was not demonstrated. In this study, the Hp fucT2 gene was successfully overexpressed in Escherichia coli. Sufficient amounts of the protein were obtained which revealed α1,2-fucosyltransferase activity to be associated with the protein. A series of substrates were chosen to examine the acceptor specificity of Hp α1,2-FucT, and the enzyme reaction products were identified by capillary electrophoresis. In contrast to the normal mammalian α1,2-FucT (H or Se enzyme), Hp α1,2-FucT prefers to use Lewis X [βGal1-4(αFuc1-3)βGlcNAc] rather than LacNAc [βGal1-4βGlcNAc] as a substrate, suggesting that H. pylori uses a novel pathway (via Lewis X) to synthesize Lewis Y. Hp α1,2-FucT also acts on type 1 acceptor [βGal1-3βGlcNAc] and Lewis a [βGal1-3(αFuc1–4)βGlcNAc], which provides H. pylori with the potential to synthesize H type 1 and Lewis b epitopes. The ability to transfer fucose to a monofucosylated substrate (Lewis X or Lewis a) makes Hp α1,2-FucT distinct from normal mammalian α1,2-FucT.
AlmR. A., LingL.-S. L., MoirD. T.20 other authors1999; Genomic sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature 397:176–180
AppelmelkB. J., Simoons-SmitI. M., NegriniR.17 other authors1996; Potential role of molecular mimicry between Helicobacter pylori lipopolysaccharide and host Lewis blood group antigens in autoimmunity. Infect Immun 64:2031–2040
AspinallG. O., MonteiroM. A.1996; Lipopolysaccharides of Helicobacter pylori strains P466 and MO19: structures of the O antigen and core oligosaccharide regions. Biochemistry 35:2498–2504[CrossRef]
AspinallG. O., MonteiroM. A., PangH., WalshE. J., MoranA. P.1996; Lipopolysaccharide of Helicobacter pylori type strain NCTC 11637 (ATCC 43504): structure of the O-antigen chain and core oligosaccharide regions. Biochemistry 35:2489–2497[CrossRef]
Blaszczyk-ThurinM., SarnestoA., ThurinI., HindsgaulO., KoprowskiH.1988; Biosynthetic pathways for the Le b and Y glycolipids in the gastric carcinoma cell line KATOIII as analyzed by a novel assay. Biochem Biophys Res Commun 151:100–108[CrossRef]
ChanN. W. C., StangierK., SherburneR., TaylorD. E., ZhangY., DovichiN., PalcicM. M.1995; The biosynthesis of Lewis X in Helicobacter pylori. . Glycobiology 5:683–688[CrossRef]
GeZ., ChanN. W. C., PalcicM. M., TaylorD. E.1997; Cloning and heterologous expression of an α1,3 fucosyltransferase gene from the gastric pathogen Helicobacter pylori. . J Biol Chem 272:21357–21363[CrossRef]
HitoshiS., KusunokiS., KanazawaI., TsujiS.1996; Molecular cloning and expression of a third type of rabbit GDP-l-fucose:β-d-galactoside 2-α-l-fucosyltransferase. J Biol Chem 271:16975–16981[CrossRef]
MartinS. L., EdbrookeM. R., HodgmanT. C., van den EijndenD. H., BirdM. I.1997; Lewis X biosynthesis in Helicobacter pylori. Molecular cloning of an α(1,3) fucosyltransferase gene. J Biol Chem 272:21349–21356[CrossRef]
MiyakeM., HakamoriS.1991; A specific cell surface glycoconjugate controlling cell motility: evidence by functional monoclonal antibodies that inhibit cell motility and tumour cell metastasis. Biochemistry 30:3328–3334[CrossRef]
MonteiroM. A., ChanK. H. N., RaskoD. A.9 other authors1998; Simultaneous expression of type 1 and type 2 Lewis blood group by Helicobacter pylori lipopolysaccharides. J Biol Chem 273:11533–11543[CrossRef]
PalcicM. M., HeerzeL. D., PierceM., HindsgaulO.1988; The use of hydrophobic synthetic glycosides as acceptors in glycosyltransferase assays. Glycoconj J 5:49–63[CrossRef]
SakamotoJ., FurukawaK., Cordon-CarloC., YinB. W. T., RettigW. J., OettgenH. F., OldL. J., LloydK. O.1986; Expression of Lewis a, Lewis b, X, and Y blood group antigens in human colonic tumours and normal tissue and in human tumour-derived cell lines. Cancer Res 46:1553–1561
SarnestoA., KohlinT., ThurinJ., Blaszczyk-ThurinM.1990; Purification of H gene-encoded β-galactoside α1-2 fucosyltransferase from human serum. J Biol Chem 265:15067–15075
SarnestoA., KohlinT., HindsgaulO., ThurinJ., Blaszczyk-ThurinM.1992; Purification of the secretor-type β-galactoside α1-2-fucosyltransferase from human serum. J Biol Chem 267:2737–2744
SunJ., ThurinJ., CooperH. S., WangP., MackiewiczM., SteplewskiZ., Blaszczyk-ThurinM.1995; Elevated expression of H type GDP-l-fucose:beta-d-galactoside alpha-2-l-fucosyltransferase is associated with human colon adenocarcinoma progression. Proc Natl Acad Sci USA 92:5724–5728[CrossRef]
TaborS., RichardsonC. C.1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA 82:1074–1078[CrossRef]
TaylorD. E., RaskoD. A., SherburneR., HoC., JewellL. D.1998; Lack of correlation between Lewis antigen expression by Helicobacter pylori and gastric epithelial cells in infected patients. Gastroenterology 115:1113–1122[CrossRef]
WangG., RaskoD. A., SherburneR., TaylorD. E.1999; Molecular genetic basis for the variable expression of Lewis Y antigen in Helicobacter pylori: Analysis of the α(1,2) fucosyltransferase gene. Mol Microbiol 31:1265–1274[CrossRef]
WatkinsW. M.1995; Biosynthesis. Molecular basis of antigenic specificity in the ABO, H, and Lewis blood group systems. In Glycoproteins pp. 313–390Edited byMontreuilJ., SchachterH., VliegenthartJ. F. G. Amsterdam: Elsevier;
WirthH.-P., YangM., KaritaM., BlaserM. J.1996; Expression of the human cell surface glycoconjugates Lewis X and Lewis Y by Helicobacter pylori isolates is related to cagA status. Infect Immun 64:4598–4605
WirthH.-P., YangM., PeekR. M., ThamK. T., BlaserM. J.1997; Helicobacter pylori Lewis expression is related to the host Lewis phenotype. Gastroenterology 113:1091–1098[CrossRef]
WirthH.-P., YangM., PeekR. M., Hook-NikanneJ., FriedM., BlaserM. J.1999; Phenotypic diversity in Lewis expression of Helicobacter pylori isolates from the same host. J Lab Clin Med 133:488–500[CrossRef]
YazawaS., NakamuraJ., AsaoT., NagamachiY., SagiM., MattaK. L., TachikawaT., AkamatsuM.1993; Aberrant α1,2 fucosyltransferases found in human colorectal carcinoma involved in the accumulation of Le b and Y antigens in colorectal tumours. Jpn J Cancer Res 84:989–995[CrossRef]