Binding of ATP by pertussis toxin and isolated toxin subunits.
The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of [3H]ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP greater than ATP greater than GTP greater than CTP greater than TTP for pertussis toxin and ATP greater than GTP greater than TTP greater than CTP for the B oligomer. Phosphate ions inhibited the binding of [3H]ATP to pertussis toxin in a competitive manner; however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of [3H]ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site.
Center for Biologics Evaluation and Research, Bethesda, Maryland 20892.
S Z
SZ
Hausman
C R
CR
Manclark
D L
DL
Burns
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001665:Binding Sites; D001667:Binding, Competitive; D009243:NAD / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D037342:Pertussis Toxin; D010710:Phosphates / Q000378:metabolism; D011485:Protein Binding; D010566:Virulence Factors, Bordetella / Q000378:metabolism*; D000509:alpha-Fetoproteins / Q000378:metabolism
D009711:Nucleotides; D010446:Peptide Fragments; D010710:Phosphates; D010566:Virulence Factors, Bordetella; D000509:alpha-Fetoproteins; D009243:NAD; D000255:Adenosine Triphosphate; D037342:Pertussis Toxin