Novel Helicobacter pylori alpha1,2-fucosyltransferase, a key enzyme in the synthesis of Lewis antigens.
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 alpha1,2-fucosyltransferase (FucT) that adds fucose to the terminal betaGal unit of the O-chain of LPS. Recently, the H. pylori (Hp) alpha1,2-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 alpha1,2-fucosyltransferase activity to be associated with the protein. A series of substrates were chosen to examine the acceptor specificity of Hp alpha1,2-FucT, and the enzyme reaction products were identified by capillary electrophoresis. In contrast to the normal mammalian alpha,2-FucT (H or Se enzyme), Hp alpha1,2-FucT prefers to use Lewis X [betaGal1-4(alphaFuc1-3)betaGlcNAc] rather than LacNAc [betaGal1-4betaGIcNAc] as a substrate, suggesting that H. pylori uses a novel pathway (via Lewis X) to synthesize Lewis Y. Hp alpha1,2-FucT also acts on type 1 acceptor [betaGal1-3betaGlcNAc] and Lewis a [betaGal1-3(alphaFuc1-4)betaGIcNAc], 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 alpha1,2-FucT distinct from normal mammalian alpha1,2-FucT.
Wang
Ge
G
Departments of Medical Microbiology and Immunology1 and Chemistry2, University of Alberta, Edmonton, AB, Canada T6G 2H7.
Boulton
Peter G
PG
Departments of Medical Microbiology and Immunology1 and Chemistry2, University of Alberta, Edmonton, AB, Canada T6G 2H7.
Chan
Nora W C
NWC
Departments of Medical Microbiology and Immunology1 and Chemistry2, University of Alberta, Edmonton, AB, Canada T6G 2H7.
Palcic
Monica M
MM
Departments of Medical Microbiology and Immunology1 and Chemistry2, University of Alberta, Edmonton, AB, Canada T6G 2H7.
Taylor
Diane E
DE
Departments of Medical Microbiology and Immunology1 and Chemistry2, University of Alberta, Edmonton, AB, Canada T6G 2H7.
D000942:Antigens, Bacterial / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D019075:Electrophoresis, Capillary; D005647:Fucosyltransferases / Q000235:genetics / Q000502:physiology*; D016480:Helicobacter pylori / Q000201:enzymology* / Q000276:immunology; D006801:Humans; D016256:Lewis X Antigen / Q000096:biosynthesis*; D014169:Transformation, Bacterial; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D005647:Fucosyltransferases; D016256:Lewis X Antigen; D000097631:Galactoside 2-alpha-L-fucosyltransferase