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Purification and characterization of the heat-labile toxin of Bordetella pertussis.

机译:百日咳博德特氏菌热不稳定毒素的纯化和鉴定。

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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.
机译:描述了从百日咳博德特氏菌细胞中纯化百日咳不耐热毒素(PEHLT)的程序。在寒冷条件下并在蛋白酶抑制剂的存在下进行的纯化程序可进行1,350倍纯化,产率约为60%。毒素显示为140 kDa的单链多肽,pI 6.02。通过在56摄氏度加热60分钟将其完全灭活。制备的针对PEHLT的兔抗血清中和了毒素,并在免疫扩散过程中产生了一条沉淀素系。在免疫扩散测定中,这种抗PEHLT血清不与百日咳毒素,丝状血凝素或百日咳腺苷酸环化酶制剂反应。纯化的PEHLT引起皮肤坏死和脾萎缩。 PEHLT非常活跃; 0.4 X 10(-12)g在新生小鼠中引起坏死性损害,对于18至20 g的小鼠,50%的致死剂量约为11 X 10(-9)g。

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