Biochemical and biological activities of recombinant S1 subunit of pertussis toxin.
degP-deficient strains of Escherichia coli grown in M-9 medium supplemented with ZnCl2 expressed the recombinant S1 subunit of pertussis toxin (rS1) in a form electrophoretically identical to the authentic S1 subunit. Subcellular fractionation showed that the full-length form of rS1 was membrane associated, while proteolytic fragments of rS1 were present in the periplasm. rS1 was extracted from outer membrane preparations with 8 M urea and purified by gel filtration chromatography. Purified rS1 ADP-ribosylated transducin at a similar molar efficiency relative to authentic pertussis toxin and, when associated with the native B oligomer of pertussis toxin, elicited Chinese hamster ovary cell clustering.
Barbieri
J T
JT
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
Pizza
M
M
Cortina
G
G
Rappuoli
R
R
D002478:Cells, Cultured; D004926:Escherichia coli / Q000032:analysis; D037342:Pertussis Toxin; D011065:Poly(ADP-ribose) Polymerases / Q000032:analysis; D011994:Recombinant Proteins / Q000032:analysis; D010566:Virulence Factors, Bordetella / Q000032:analysis* / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D011065:Poly(ADP-ribose) Polymerases; D037342:Pertussis Toxin; D015032:Zinc