Expression and immunological properties of the five subunits of pertussis toxin.
Pertussis toxin, a protein composed of five different subunits, is responsible for the pathogenicity of Bordetella pertussis and is the main component of a new vaccine against whooping cough. The genes coding for the five subunits, recently cloned and sequenced, are organized as an operon. We approached the problem of expression of the five genes in Escherichia coli and, although we obtained high levels of transcription of the native pertussis toxin genes, the amount of proteins produced was very low or undetectable. To obtain suitable expression of each of the five subunits, we fused their genes to the gene coding for the DNA polymerase of MS2 in the expression vector pEx31. A total of 5 to 30 mg of purified fusion proteins could be obtained from 1 liter of culture. The purified fusion proteins were used to immunize rabbits to obtain sera against each of the five subunits. These sera, although able to recognize the toxin in an enzyme-linked immunosorbent assay and the corresponding subunits in Western blots, were not able to protect CHO cells from the action of pertussis toxin. Mice immunized with the five subunits were not protected from an intracerebral challenge with B. pertussis. Subunits S2 and S3, which are 67% homologous, were shown to cross-react immunologically. The fused subunit S1 was able to ADP-ribosylate transducin as efficiently as the native pertussis toxin.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli; D005798:Genes, Bacterial; D046911:Macromolecular Substances; D051379:Mice; D037342:Pertussis Toxin; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000235:genetics* / Q000276:immunology
D001428:Bacterial Vaccines; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; D037342:Pertussis Toxin