Identification and characterization of binding properties of Helicobacter pylori by glycoconjugate arrays.
The microaerophilic bacterium Helicobacter pylori is well established for its role in development of different gastric diseases. Bacterial adhesins and corresponding binding sites on the epithelial surface allow H. pylori to colonize the gastric tissue. In this investigation, the adhesion of H. pylori to dot blot arrays of natural glycoproteins and neoglycoproteins was studied. Adhesion was detected by overlay with fluorescence-labeled bacteria on immobilized (neo)glycoproteins. The results confirmed the interaction between the adhesin BabA and the H-1-, Lewis b-, and related fucose-containing antigens. In addition, H. pylori bound to terminal alpha2-3-linked sialic acids as previously described. The use of a sabA mutant and sialidase treatment of glycoconjugate arrays showed that the adherence of H. pylori to laminin is mediated by the sialic acid-binding adhesin, SabA. The adhesion to salivary mucin MUC5B is mainly associated with the BabA adhesin and to a lesser extent with the SabA adhesin. This agrees with reports, that MUC5B carries both fucosylated blood group antigens and alpha2-3-linked sialic acids. The adhesion of H. pylori to fibronectin and lactoferrin persisted in the babA/sabA double mutant. Because binding to these molecules was abolished by denaturation rather than by deglycosylation, it was suggested to depend on the recognition of unknown receptor moieties by an additional unknown bacterial surface component. The results demonstrate that the bacterial overlay method on glycoconjugate arrays is a useful tool for exploration and the characterization of unknown adhesin specificities of H. pylori and other bacteria.
Walz
Anke
A
Department of Operative Dentistry and Periodontology, Dental School, University of Regensburg, D-93053 Regensburg, Germany.
Odenbreit
Stefan
S
Mahdavi
Jafar
J
Borén
Thomas
T
Ruhl
Stefan
S
D018829:Adhesins, Bacterial / Q000378:metabolism; D001422:Bacterial Adhesion; D002240:Carbohydrate Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism; D006001:Glycoconjugates / Q000737:chemistry / Q000378:metabolism*; D016480:Helicobacter pylori / Q000235:genetics / Q000502:physiology*; D007781:Lactoferrin / Q000378:metabolism; D008969:Molecular Sequence Data
D018829:Adhesins, Bacterial; C400429:BabA protein, Helicobacter pylori; D005353:Fibronectins; D006001:Glycoconjugates; C500808:SabA protein, Helicobacter pylori; D007781:Lactoferrin