Properties of pertussis toxin B oligomer assembled in vitro from recombinant polypeptides produced by Escherichia coli.
The subunits that make up the pentameric B oligomer of pertussis toxin (S2, S3, S4, and S5) were individually synthesized as recombinant polypeptides in Escherichia coli, isolated as insoluble inclusion bodies, and assembled into a multimeric form in vitro by spontaneous association following treatment with a chaotropic agent, reduction, and reoxidation. The recombinant B multimer, purified by fetuin-Sepharose affinity chromatography, contained all four of the individual subunits and possessed the mitogenic and hemagglutinating activities characteristic of the native B oligomer. Immunization of mice with the recombinant B oligomer elicited antibodies that neutralized pertussis toxin in vitro and, moreover, provided protection in vivo against the leukocytosis-promoting activity of the toxin. These results demonstrate the potential for assembly of complex multimeric proteins from recombinant DNA-derived polypeptides and provide a novel means for production of an acellular pertussis vaccine component.
Burnette
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Amgen Inc., Amgen Center, Thousand Oaks, California 91320-1789.
Arciniega
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JL
Mar
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Burns
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D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000378:metabolism*; D007114:Immunization; D051379:Mice; D037342:Pertussis Toxin; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis / Q000276:immunology*
D000907:Antibodies, Bacterial; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin