1887

Abstract

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 (Hp) α1,2-FucT-encoding gene () 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 gene was successfully overexpressed in . 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 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 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.

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1999-11-01
2020-01-21
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References

  1. Alm R. A., Ling L.-S. L., Moir D. T..20 other authors 1999; Genomic sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature397:176–180
    [Google Scholar]
  2. Appelmelk B. J., Simoons-Smit I. M., Negrini R..17 other authors 1996; Potential role of molecular mimicry between Helicobacter pylori lipopolysaccharide and host Lewis blood group antigens in autoimmunity. Infect Immun64:2031–2040
    [Google Scholar]
  3. Appelmelk B. J., Shiberu B., Trinks C..10 other authors 1998; Phase variation in Helicobacter pylori lipopolysaccharide. Infect Immun66:70–76
    [Google Scholar]
  4. Aspinall G. O., Monteiro M. A.. 1996; Lipopolysaccharides of Helicobacter pylori strains P466 and MO19: structures of the O antigen and core oligosaccharide regions. Biochemistry35:2498–2504[CrossRef]
    [Google Scholar]
  5. Aspinall G. O., Monteiro M. A., Pang H., Walsh E. J., Moran A. P.. 1996; Lipopolysaccharide of Helicobacter pylori type strain NCTC 11637 (ATCC 43504): structure of the O-antigen chain and core oligosaccharide regions. Biochemistry35:2489–2497[CrossRef]
    [Google Scholar]
  6. Avent N. D.. 1997; Human erythrocyte antigen expression its molecular bases. Brit J Biomed Sci54:16–37
    [Google Scholar]
  7. Blaszczyk-Thurin M., Sarnesto A., Thurin I., Hindsgaul O., Koprowski H.. 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 Commun151:100–108[CrossRef]
    [Google Scholar]
  8. Chan N. W. C., Stangier K., Sherburne R., Taylor D. E., Zhang Y., Dovichi N., Palcic M. M.. 1995; The biosynthesis of Lewis X in Helicobacter pylori. . Glycobiology5:683–688[CrossRef]
    [Google Scholar]
  9. Ge Z., Chan N. W. C., Palcic M. M., Taylor D. E.. 1997; Cloning and heterologous expression of an α1,3 fucosyltransferase gene from the gastric pathogen Helicobacter pylori. . J Biol Chem272:21357–21363[CrossRef]
    [Google Scholar]
  10. Hakomori S.-I.. 1989; Aberrant glycosylation in tumours and tumour-associated carbohydrate antigens. Adv Cancer Res52:257–331
    [Google Scholar]
  11. Herry S., Oriol R., Samuelsson B.. 1995; Lewis histo-blood group system and associated secretory phenotypes. Vox Sang69:166–182[CrossRef]
    [Google Scholar]
  12. Hitoshi S., Kusunoki S., Kanazawa I., Tsuji S.. 1996; Molecular cloning and expression of a third type of rabbit GDP-l-fucose:β-d-galactoside 2-α-l-fucosyltransferase. J Biol Chem271:16975–16981[CrossRef]
    [Google Scholar]
  13. Kannagi R.. 1997; Carbohydrate-mediated cell adhesion involved in hematogenous metastasis of cancer. Glycoconj J14:577–584[CrossRef]
    [Google Scholar]
  14. Kleene R., Berger E. G.. 1993; The molecular and cell biology of glycosyltransferases. Biochim Biophys Acta1154:283–325[CrossRef]
    [Google Scholar]
  15. Martin S. L., Edbrooke M. R., Hodgman T. C., van den Eijnden D. H., Bird M. I.. 1997; Lewis X biosynthesis in Helicobacter pylori. Molecular cloning of an α(1,3) fucosyltransferase gene. J Biol Chem272:21349–21356[CrossRef]
    [Google Scholar]
  16. Miyake M., Hakamori S.. 1991; A specific cell surface glycoconjugate controlling cell motility: evidence by functional monoclonal antibodies that inhibit cell motility and tumour cell metastasis. Biochemistry30:3328–3334[CrossRef]
    [Google Scholar]
  17. Monteiro M. A., Chan K. H. N., Rasko D. A..9 other authors 1998; Simultaneous expression of type 1 and type 2 Lewis blood group by Helicobacter pylori lipopolysaccharides. J Biol Chem273:11533–11543[CrossRef]
    [Google Scholar]
  18. Palcic M. M., Heerze L. D., Pierce M., Hindsgaul O.. 1988; The use of hydrophobic synthetic glycosides as acceptors in glycosyltransferase assays. Glycoconj J5:49–63[CrossRef]
    [Google Scholar]
  19. Sakamoto J., Furukawa K., Cordon-Carlo C., Yin B. W. T., Rettig W. J., Oettgen H. F., Old L. J., Lloyd K. 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 Res46:1553–1561
    [Google Scholar]
  20. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  21. Sarnesto A., Kohlin T., Thurin J., Blaszczyk-Thurin M.. 1990; Purification of H gene-encoded β-galactoside α1-2 fucosyltransferase from human serum. J Biol Chem265:15067–15075
    [Google Scholar]
  22. Sarnesto A., Kohlin T., Hindsgaul O., Thurin J., Blaszczyk-Thurin M.. 1992; Purification of the secretor-type β-galactoside α1-2-fucosyltransferase from human serum. J Biol Chem267:2737–2744
    [Google Scholar]
  23. Sherburne R., Taylor D. E.. 1995; Helicobacter pylori expresses a complex surface carbohydrate, Lewis X.. Infect Immun63:4564–4568
    [Google Scholar]
  24. Sun J., Thurin J., Cooper H. S., Wang P., Mackiewicz M., Steplewski Z., Blaszczyk-Thurin M.. 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 USA92:5724–5728[CrossRef]
    [Google Scholar]
  25. Tabor S., Richardson C. C.. 1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA82:1074–1078[CrossRef]
    [Google Scholar]
  26. Taylor D. E., Rasko D. A., Sherburne R., Ho C., Jewell L. D.. 1998; Lack of correlation between Lewis antigen expression by Helicobacter pylori and gastric epithelial cells in infected patients. Gastroenterology115:1113–1122[CrossRef]
    [Google Scholar]
  27. Tomb J.-F., White O., Kerlavage A. R..39 other authors 1997; The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature388:539–547
    [Google Scholar]
  28. Wang G., Rasko D. A., Sherburne R., Taylor D. 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 Microbiol31:1265–1274[CrossRef]
    [Google Scholar]
  29. Watkins W. M.. 1995; Biosynthesis. Molecular basis of antigenic specificity in the ABO, H, and Lewis blood group systems. In Glycoproteins pp.313–390Edited by Montreuil J., Schachter H., Vliegenthart J. F. G.. Amsterdam: Elsevier;
    [Google Scholar]
  30. Wirth H.-P., Yang M., Karita M., Blaser M. J.. 1996; Expression of the human cell surface glycoconjugates Lewis X and Lewis Y by Helicobacter pylori isolates is related to cagA status. Infect Immun64:4598–4605
    [Google Scholar]
  31. Wirth H.-P., Yang M., Peek R. M., Tham K. T., Blaser M. J.. 1997; Helicobacter pylori Lewis expression is related to the host Lewis phenotype. Gastroenterology113:1091–1098[CrossRef]
    [Google Scholar]
  32. Wirth H.-P., Yang M., Peek R. M., Hook-Nikanne J., Fried M., Blaser M. J.. 1999; Phenotypic diversity in Lewis expression of Helicobacter pylori isolates from the same host. J Lab Clin Med133:488–500[CrossRef]
    [Google Scholar]
  33. Yao Z., Liu H., Valvano M. A.. 1992; Acetylation of O-specific lipopolysaccharides from Shigella flexneri 3a and 2a occurs in Escherichia coli K-12 carrying cloned S. flexneri 3a and 2a rfb genes. J Bacteriol174:7500–7508
    [Google Scholar]
  34. Yazawa S., Nakamura J., Asao T., Nagamachi Y., Sagi M., Matta K. L., Tachikawa T., Akamatsu M.. 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 Res84:989–995[CrossRef]
    [Google Scholar]
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