Human gastric glycosphingolipids recognized by Helicobacter pylori vacuolating cytotoxin VacA.
Many bacterial toxins utilize cell surface glycoconjugate receptors for attachment to target cells. In the present study the potential carbohydrate binding of Helicobacter pylori vacuolating cytotoxin VacA was investigated by binding to human gastric glycosphingolipids on thin-layer chromatograms. Thereby a distinct binding of the toxin to two compounds in the non-acid glycosphingolipid fraction was detected. The VacA-binding glycosphingolipids were isolated and characterized by mass spectrometry and proton NMR as galactosylceramide (Galbeta1Cer) and galabiosylceramide (Galalpha4Galbeta1Cer). Comparison of the binding preferences of the protein to reference glycosphingolipids from other sources showed an additional recognition of glucosylceramide (Glcbeta1Cer), lactosylceramide (Galbeta4Glcbeta1Cer) and globotriaosylceramide (Galalpha4Galbeta4Glcbeta1Cer). No binding to the glycosphingolipids recognized by the VacA holotoxin was obtained with a mutant toxin with deletion of the 37 kDa fragment of VacA (P58 molecule). Collectively our data show that the VacA cytotoxin is a glycosphingolipid binding protein, where the 37 kDa moiety is required for carbohydrate recognition. The ability to bind to short chain glycosphingolipids will position the toxin close to the cell membrane, which may facilitate toxin internalization.
Roche
Niamh
N
Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Göteborg University, PO Box 440, S-405 30 Göteborg, Sweden.
Ilver
Dag
D
Angström
Jonas
J
Barone
Silvia
S
Telford
John L
JL
Teneberg
Susann
S
D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D004848:Epithelium / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism*; D016481:Helicobacter Infections / Q000276:immunology; D016480:Helicobacter pylori / Q000737:chemistry*; D006801:Humans; D013270:Stomach / Q000737:chemistry*
D001426:Bacterial Proteins; D006028:Glycosphingolipids; C074689:VacA protein, Helicobacter pylori