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Microbiology 145 (1999), 3245-3253
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Microbiology (1999), 145, 3245-3253.
© 1999 Society for General Microbiology

Biochemistry

Novel Helicobacter pylori {alpha}1,2-fucosyltransferase, a key enzyme in the synthesis of Lewis antigens

Ge Wang1, Peter G. Boulton1, Nora W. C. Chan2, Monica M. Palcic2 and Diane E. Taylor1

Departments of Medical Microbiology and Immunology1 and Chemistry2, University of Alberta, Edmonton, AB, Canada T6G 2H7

Author for correspondence: Diane E. Taylor. Tel: +1 780 492 4777. Fax: +1 780 492 7521. e-mail: diane.taylor{at}ualberta.ca

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 {alpha}1,2-fucosyltransferase (FucT) that adds fucose to the terminal ßGal unit of the O-chain of LPS. Recently, the H. pylori (Hp) {alpha}1,2-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 {alpha}1,2-fucosyltransferase activity to be associated with the protein. A series of substrates were chosen to examine the acceptor specificity of Hp {alpha}1,2-FucT, and the enzyme reaction products were identified by capillary electrophoresis. In contrast to the normal mammalian {alpha}1,2-FucT (H or Se enzyme), Hp {alpha}1,2-FucT prefers to use Lewis X [ßGal1-4({alpha}Fuc1-3)ßGlcNAc] rather than LacNAc [ßGal1-4ßGlcNAc] as a substrate, suggesting that H. pylori uses a novel pathway (via Lewis X) to synthesize Lewis Y. Hp {alpha}1,2-FucT also acts on type 1 acceptor [ßGal1-3ßGlcNAc] and Lewis a [ßGal1-3({alpha}Fuc1–4)ßGlcNAc], 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 {alpha}1,2-FucT distinct from normal mammalian {alpha}1,2-FucT.

Keywords: Helicobacter pylori, {alpha}1,2-fucosyltransferase, Lewis antigens

Abbreviations: FucT, fucosyltransferase; Hp, H. pylori; LacNAc, N-acetyllactosamine; Lea, Lewis a; Leb, Lewis b; LeX, Lewis X; LeY, Lewis Y; TMR, tetramethylrhodamine




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