Abstract
The human gastric pathogen Helicobacter pylori has been shown to bind to glycoconjugates of human leucocytes in a sialic acid-dependent way. In order to improve the identification of the binding epitope, a new technique was developed to analyze the ketosidic linkage position between a terminal sialic acid and the consecutive monosaccharide. Permethylation and reduction with LiAlH4 followed by trifluoroacetolysis in 1000:1 trifluoroacetic anhydride:trifluoroacetic acid (24 h, 100 °C) results in the cleavage of glycosidic but not ketosidic bonds. The disaccharide products were analyzed by gas chromatography-mass spectrometry and sialyl-3 or -6 position and NeuAc or NeuGc are identified by their separate retention times and mass spectra. The method was worked out on model saccharides and applied on five-sugar gangliosides (sialylparaglobosides) of human leucocytes. Radiolabeled Helicobacter pylori was shown to bind to the upper part, but not to the lower part, of the five-sugar interval of a mixture of gangliosides separated on a thin-layer chromatogram. Using a membrane blotting procedure the active and inactive bands were isolated and shown to be NeuAcα2-3- and NeuAcα2-6-paraglobosides, respectively.
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References
Varki A (1992) Glycobiology 2: 25-40.
Rosenberg A (1995) (ed) Biology of the Sialic Acids. New York: Plenum Press, 1995.
Varki A (1993) Glycobiology 3: 97-130.
Karlsson K-A (1995) Curr Opin Struct Biol 5: 622-35.
Wadström T (1995) Curr Opin Gastroenterol 11: 69-75.
Evans DG, Evans DJ, Moulds JJ, Graham DY (1988) Infect Immun 56: 2896-906.
Hirmo S, Kelm S, Schauer R, Nilsson B, Wadström T (1996) Glycoconjugate J 13: 1005-11.
Miller-Podraza H, Abul Milh M, Bergström J, Karlsson K-A (1996) Glycoconjugate J 13: 453-60.
Miller-Podraza H, Bergström J, Abul Milh M, Karlsson K-A (1997) Glycoconjugate J 14: 467-71.
Karlsson H, Carlstedt I, Hansson GC (1989) Anal Biochem 182: 438-46.
Uchida Y, Tsukada Y, Sugimori T (1979) J Biochem 86: 1573-85.
Gunnarsson A (1984) Trifluoroacetolysis, Specific techniques for the isolation of biologically active oligosaccharides from glycoconjugates Ph.D. Thesis, University of Lund, Sweden.
Karlsson K-A (1987) Methods Enzymol 138: 212-20.
Ciucanu I, Kerek F (1984) Carb Res 131: 209-17.
Larson G, Karlsson H, Hansson GC, Pimlott W (1987) Carb Res 161: 281-90.
Karlsson K-A (1974) Biochemistry 13: 3643-7.
Taki T, Handa S, Ishikawa D(1994) Anal Biochem 221: 312-16.
Karlsson K-A, Strömberg N (1987) Methods Enzymol 138: 220-32.
Miller-Podraza H, Stenhagen G, Larsson T, Andersson C, Karlsson K-A (1997) Glycoconjugate J 14: 231-9.
Waldi D (1962) In Dünnschicht Chromatographie. Ein Laboratoriumshandbuch (Stahl E, ed) pp 496-515. Berlin: Springer-Verlag.
Stroud MR, Handa K, Ito K, Salyan MEK, Fang H, Levery SB, Hakomori S, Reinhold BB, Reinhold VN (1995) Biochem Biophys Res Com 209: 777-87.
Karlsson K-A (1973) FEBS Letters 32: 317-20.
Karlsson K-A (1976) In Glycolipid Methodology (Witting LA, ed) pp 97-122. Champaign, Illinois: American Oil Chemists' Society.
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Johansson, L., Karlsson, KA. Selective binding by Helicobacter pylori of leucocyte gangliosides with 3-linked sialic acid, as identified by a new approach of linkage analysis. Glycoconj J 15, 713–721 (1998). https://doi.org/10.1023/A:1006992616254
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DOI: https://doi.org/10.1023/A:1006992616254
- Helicobacter pylori
- leucocytes
- glycosphingolipids
- trifluoroacetolysis
- sialic acid
- gas chromatography-mass spectrometry
- linkage position
- NeuAc, N-acetylneuraminic acid
- NeuGc, N-glycolylneuraminic acid
- Gal, galactose
- GlcNAc, N-acetylglucosamine
- GC/MS, gas chromatography mass spectrometry
- DMSO, dimethyl sulphoxide
- ether, diethyl ether
- TFA, trifluoroacetic acid
- TFAA, trifluoroacetic anhydride
- TFAc, trifluoroacetyl
- MALDI-TOF MS, matrix assisted laser desorptionionization time of flight mass spectrometry
- 6-SL, 6-sialyllactose
- 3-SL, 3-sialyllactose
- SPG, sialylparagloboside
- Me, methyl
- FAB MS, fast atom bombardment mass spectrometry
- EI MS, electron ionization mass spectrometry
- DHB, 2,5-dihydroxybenzoic acid
- TLC, thin-layer chromatography