Binding of pertussis toxin to eucaryotic cells and glycoproteins.
The binding of pertussis toxin and its subunits to cell surface receptors and purified glycoproteins was examined. The interaction of pertussis toxin with components of two variant Chinese hamster ovary (CHO) cell lines was studied. These cell lines are deficient in either sialic acid residues (LEC 2) or sialic acid and galactose residues (LEC 8) on cell surface macromolecules. The binding of pertussis toxin to components of these cells differed from the binding of the toxin to wild-type components. Although the toxin bound to a 165,000-dalton glycoprotein found in N-octylglucoside extracts of wild-type cells, it did not bind to components found in extracts of LEC 2 cells. In contrast, the toxin bound to components found in extracts of LEC 8 cells, which are variant cells that contain increased amounts of terminal N-acetylglucosamine residues on cell surface macromolecules. These results suggest that the receptor for pertussis toxin on CHO cells contains terminal acetamido-containing sugars. The cytopathic effect of the toxin on both types of variant cells was much reduced compared with its effects on wild-type cells. Thus, optimal functional binding of pertussis toxin appears to require a complete sialyllactosamine (NeuAc----Gal beta 4GlcNAc) sequence on surface macromolecules. In addition to studying the nature of the eucaryotic receptor for pertussis toxin, we examined corresponding binding sites for glycoproteins on the toxin molecule. Binding of both S2-S4 and S3-S4 dimers of the toxin to cellular components and purified glycoproteins was observed. The two dimers bound to a number of glycoproteins containing N-linked oligosaccharides but not O-linked oligosaccharides, and differences in the binding of the two dimers to some glycoproteins was noted. These data indicate that the holotoxin molecule contains at least two glycoprotein-binding sites which may have slightly different specificities for glycoproteins.
Witvliet
M H
MH
Laboratory of Pertussis, Food and Drug Administration, Bethesda, Maryland 20892.
Burns
D L
DL
Brennan
M J
MJ
Poolman
J T
JT
Manclark
C R
CR
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D002460:Cell Line; D006224:Cricetinae; D004910:Erythrocyte Membrane / Q000378:metabolism; D005057:Eukaryotic Cells / Q000378:metabolism; D005777:Geese / Q000097:blood; D006023:Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D037342:Pertussis Toxin; D010566:Virulence Factors, Bordetella / Q000378:metabolism*; D000509:alpha-Fetoproteins / Q000378:metabolism
D006023:Glycoproteins; D010566:Virulence Factors, Bordetella; D000509:alpha-Fetoproteins; D000246:Adenosine Diphosphate Ribose; D037342:Pertussis Toxin