Sulfated glycoconjugate receptors for the Bordetella pertussis adhesin filamentous hemagglutinin (FHA) and mapping of the heparin-binding domain on FHA.
Filamentous hemagglutinin (FHA) is a major adhesin present on the surface of the gram-negative respiratory pathogen Bordetella pertussis. A number of binding mechanisms have been described for the interaction of FHA with eukaryotic cells. We have focused on its function as a sulfated polysaccharide-binding protein and on identifying potential receptors for FHA on the epithelial cell surface. Using a thin-layer overlay technique, we found that FHA binds specifically to sulfated glycolipids but not to gangliosides or other neutral glycolipids. These results suggest that epithelial cell surface sulfated glycolipids function as receptors for FHA. Further studies demonstrated that a Chinese hamster ovary (CHO) cell strain deficient in glycosaminoglycan expression exhibits greatly diminished attachment to FHA. By FHA-Affi-Gel chromatography, a putative receptor for FHA that has characteristics consistent with a heparan sulfate proteoglycan was isolated from epithelial cell extracts. In addition, by using recombinant FHA fusion proteins, a specific glycosaminoglycan-binding domain located near the N terminus of the FHA molecule was identified. Our results indicate that the B. pertussis adhesin FHA may utilize sulfated glycolipids and proteoglycans commonly found on the surface of human cells and tissues to initiate infection.
Hannah
J H
JH
Division of Bacterial Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892.
Menozzi
F D
FD
Renauld
G
G
Locht
C
C
Brennan
M J
MJ
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001884:Bordetella; D001886:Bordetella pertussis / Q000472:pathogenicity*; D016466:CHO Cells; D006224:Cricetinae; D017931:DNA Primers / Q000737:chemistry; D005798:Genes, Bacterial; D006017:Glycolipids / Q000378:metabolism; D006367:HeLa Cells; D006388:Hemagglutinins / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015183:Restriction Mapping; D013433:Sulfoglycosphingolipids / Q000378:metabolism; D010566:Virulence Factors, Bordetella
D018829:Adhesins, Bacterial; D017931:DNA Primers; D006017:Glycolipids; D006388:Hemagglutinins; D006497:Heparan Sulfate; D013433:Sulfoglycosphingolipids; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis