Structural characterization of pertussis toxin A subunit.
The relationship between the structure of the A subunit of pertussis toxin and its function was analyzed. Limited tryptic digestion of the A subunit converted the protein to two stable fragments (Mr = 20,000 and 18,000). Antibodies raised to synthetic peptides homologous to regions in the A subunit were used to map these fragments. Both fragments were shown to contain the NH2-terminal portion but not the COOH-terminal portion of the A subunit. While these fragments exhibited NAD glycohydrolase activity, they were unable to reassociate with the B oligomer of the toxin. Thus the COOH-terminal portion of the A subunit does not contain the residues which are required for the NAD glycohydrolase activity of the toxin. However, this region of the molecule may be important for maintaining the oligomeric structure of the toxin. These results suggest that the A subunit of pertussis toxin is similar in structure to the A subunit of cholera toxin. In addition, antibodies raised to a synthetic peptide identical to residues 6-17 of the A subunit of pertussis toxin will bind to the A subunit of cholera toxin.
Burns
D L
DL
Center for Drugs and Biologics, Food and Drug Administration, Bethesda, Maryland 20892.
Hausman
S Z
SZ
Lindner
W
W
Robey
F A
FA
Manclark
C R
CR
D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D002772:Cholera Toxin; D007163:Immunosorbent Techniques; D051379:Mice; D008970:Molecular Weight; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D009244:NAD+ Nucleosidase; D010446:Peptide Fragments / Q000378:metabolism; D010449:Peptide Mapping; D037342:Pertussis Toxin; D011487:Protein Conformation; D014357:Trypsin / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000378:metabolism
D000906:Antibodies; D010446:Peptide Fragments; D010566:Virulence Factors, Bordetella; D002772:Cholera Toxin; D037342:Pertussis Toxin; D009699:N-Glycosyl Hydrolases; D009244:NAD+ Nucleosidase; D014357:Trypsin