Activities of complete and truncated forms of pertussis toxin subunits S1 and S2 synthesized by Escherichia coli.
The genes encoding the S1 and S2 subunits of pertussis toxin were expressed in Escherichia coli under lac operon transcription and translation control with pUC8 and pUC18 as the expression vectors. Various versions of the subunits were detected with anti-S1 or anti-S2 monoclonal antibodies. Recombinant S1, but not S2, subunit contained the enzymatic NAD-glycohydrolase and NAD:Gi ADP-ribosyltransferase activities. Both activities were also expressed by a truncated version of the S1 subunit in which the 48 carboxy-terminal amino acid residues, including a predicted Rossman structure and one of the two cysteines, had been deleted. The epitope for an anti-S2 monoclonal antibody was localized to the N-terminal 40-amino-acid region of the S2 subunit. Both the S1 and S2 subunits expressed in E. coli reacted with human hyperimmune serum. The full length and the truncated recombinant S1 subunit also reacted in Western blots with a neutralizing and protective monoclonal anti-S1 antibody. The different versions of S1 and S2 subunits expressed in E. coli are useful for mapping active sites, epitopes, and regions that interact with receptors or the other subunits in the holotoxin. These recombinant subunits will also facilitate the development of a safer, new-generation vaccine against whooping cough.
D036002:ADP Ribose Transferases; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000941:Antigens / Q000235:genetics*; D001428:Bacterial Vaccines / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli; D019204:GTP-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D010430:Pentosyltransferases / Q000235:genetics; D037342:Pertussis Toxin; D014614:Vaccines, Synthetic / Q000235:genetics*; D010566:Virulence Factors, Bordetella / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000907:Antibodies, Bacterial; D000941:Antigens; D001428:Bacterial Vaccines; D046911:Macromolecular Substances; D014614:Vaccines, Synthetic; D010566:Virulence Factors, Bordetella; D036002:ADP Ribose Transferases; D010430:Pentosyltransferases; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins