Interaction of Helicobacter pylori with sialylated carbohydrates: the dependence on different parts of the binding trisaccharide Neu5Ac{alpha}3Gal{beta}4GlcNAc.
We have recently shown that binding of Helicobacter pylori to sialylated carbohydrates is dependent on the presence of the carboxyl group and the glycerol chain of Neu5Ac. In this work we investigated the importance of GlcNAc in the binding trisaccharide Neu5Acalpha3Galbeta4GlcNAc and the role of the N-acetamido groups of both Neu5Ac and GlcNAc. An important part of the project was epitope dissection, that is chemical derivatizations of the active carbohydrate followed by binding studies. In addition we used a panel of various unmodified carbohydrate structures in the form of free oligosaccharides or glycolipids. These were tested for binding by hemagglutination inhibition assay, TLC overlay tests, and a new quantitative approach using radiolabeled neoglycoproteins. The studies showed that the N-acetamido group of Neu5Ac is important for binding by H. pylori, whereas the same group of GlcNAc is not. In addition, Fuc attached to GlcNAc, as tested with sialyl-Lewis x, did not affect the binding. Free Neu5Ac was inactive as inhibitor, and Neu5Acalpha3Gal turned out to be active. The binding preference for neolacto structures was confirmed, although one strain also was inhibited by lacto chains. The combined results revealed that an intact Neu5Ac is crucial for the interactions with H. pylori. Parts of Gal also seem to be necessary, whereas the role of the GlcNAc is secondary. GlcNAc does influence binding, however, primarily serving as a guiding carrier for the binding epitope rather than being a part of the binding structure.
Johansson
Petra
P
Institute of Medical Biochemistry, Göteborg University, PO Box 440, SE 405 30 Göteborg, Sweden. petra.johansson@medkem.gu.se
Nilsson
Jonas
J
Angström
Jonas
J
Miller-Podraza
Halina
H
D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D018360:Crystallography, X-Ray; D005915:Globosides / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry; D016480:Helicobacter pylori / Q000737:chemistry* / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests / Q000379:methods; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D014312:Trisaccharides / Q000737:chemistry* / Q000378:metabolism
D002241:Carbohydrates; D005915:Globosides; D006023:Glycoproteins; D014312:Trisaccharides