Purification and characterization of the heat-labile toxin of Bordetella pertussis.
A procedure is described for purification of pertussis heat-labile toxin (PEHLT) from cells of Bordetella pertussis. The purification procedure, performed in the cold and in the presence of protease inhibitors, gives 1,350-fold purification with yields of about 60%. The toxin was shown to be a single-chain polypeptide of 140 kDa, pI 6.02. It was completely inactivated by heating at 56 degrees C for 60 min. Rabbit antiserum prepared against PEHLT neutralized the toxin and gave a single precipitin line on immunodiffusion. In immunodiffusion assays, this anti-PEHLT serum did not react with pertussis toxin, filamentous hemagglutinin, or preparations of pertussis adenylate cyclase. Purified PEHLT elicited dermonecrosis and atrophy of the spleen. PEHLT is extraordinarily active; 0.4 X 10(-12) g caused necrotic lesions in newborn mice, and with 18- to 20-g mice the 50% lethal dose was about 11 X 10(-9) g.
Zhang
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YL
Laboratory of Molecular and Developmental Immunity, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
Sekura
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RD
D000262:Adenylyl Cyclases / Q000302:isolation & purification; D000818:Animals; D001427:Bacterial Toxins / Q000276:immunology / Q000302:isolation & purification* / Q000633:toxicity; D001886:Bordetella pertussis / Q000378:metabolism* / Q000472:pathogenicity; D006358:Hot Temperature; D051379:Mice; D008970:Molecular Weight; D009500:Neutralization Tests; D011817:Rabbits; D011503:Transglutaminases; D010566:Virulence Factors, Bordetella
D001427:Bacterial Toxins; D010566:Virulence Factors, Bordetella; C060969:dermonecrotic toxin, Bordetella; D011503:Transglutaminases; D000262:Adenylyl Cyclases