Gureeva A A, Grigorian G Iu, Rybin V O, Pereverzev N A, Lapaeva I A
Zh Mikrobiol Epidemiol Immunobiol. 1986 Jul(7):32-7.
The modified method for the isolation and purification of B. pertussis toxin has been proposed. Chromatography with the use of hydroxylapatite and lentil lectin--Sepharose 4B has permitted the isolation of the preparation purified 600 times. Its molecular weight is about 90,000. The preparation has been found to possess leukocytosis-stimulating, histamine-sensitising and hemagglutinating activity. Electrophoretic analysis has revealed that the isolated substance consists of four subunits with molecular weights 28,400, 24,300, 21,800 and 15,200. This substance has proved to be capable of hydrolyzing NAD+, as well as of suppressing the GTPase activity of transducin, which is indicative of the covalent modification (ADP-ribosylyzing) of GTP-binding protein under the action of B. pertussis toxin. Two methods for the isolation of B. pertussis toxin (from liquid and solid growth media), as well as the isolation of the toxin from different B. pertussis strains, are evaluated.
已提出百日咳博德特氏菌毒素分离纯化的改良方法。使用羟基磷灰石和扁豆凝集素 - 琼脂糖4B进行色谱分离，已能够分离出纯化了600倍的制剂。其分子量约为90,000。已发现该制剂具有刺激白细胞增多、组胺致敏和血凝活性。电泳分析表明，分离出的物质由分子量分别为28,400、24,300、21,800和15,200的四个亚基组成。已证明该物质能够水解NAD +，并抑制转导蛋白的GTP酶活性，这表明在百日咳博德特氏菌毒素作用下，GTP结合蛋白发生了共价修饰（ADP - 核糖基化）。对两种从百日咳博德特氏菌毒素中分离毒素的方法（从液体和固体生长培养基中分离）以及从不同百日咳博德特氏菌菌株中分离毒素的方法进行了评估。
