Separation of toxin and hemagglutinin from crystalline toxin of Clostridium botulinum type A by anion exchange chromatography and determination of their dimensions by gel filtration.
Separation of toxin and hemagglutinin from crystalline toxin of Clostridium botulinum type A by anion exchange chromatography and determination of their dimensions by gel filtration.
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通过阴离子交换色谱法从 A 型肉毒杆菌结晶毒素中分离毒素和血凝素，并通过凝胶过滤测定它们的尺寸。
DOI：
10.1016/s0021-9258(18)93623-9
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发表时间：
1968
期刊：
影响因子：
--
通讯作者：
D. A. Boroff
中科院分区：
文献类型：
--
作者：
B. Dasgupta;D. A. Boroff
A型肉毒梭菌毒素的结晶制剂在DEAE-纤维素柱上，在pH 8.0的Tris-HCl缓冲液中进行分级。分离得到的毒素组分无血凝活性，比活性为晶体毒素的5倍。第二组分是一种强大的血凝素，但毒性很弱；在不同的洗脱条件下，它从柱子中以一个或多个峰的形式出现。通过重层析和免疫学试验，证明第二组分的毒性是由于被微量的有毒组分污染所致。根据几个标准，毒性和血凝成分似乎至少是两种不同的蛋白质。从这些物质的超速离心分析中获得的大部分信息也是通过Sephadex G-200柱凝胶过滤获得的，蛋白质浓度要小得多。在生理pH下，毒性组分的相对分子质量为15万，斯托克斯半径为48A；这些尺寸与其他研究人员在活体中建立的尺寸一致。血凝素呈现三种聚集态，分子量分别为29万、50万和90万。
Crystalline preparations ofClostridium botulinumtype A toxin were fractionated on DEAE-cellulose columns with Tris-HCl buffers at pH 8.0. The isolated toxic fraction was free of hemagglutinating activity and contained 5 times the specific activity of the crystalline toxin. The second fraction was a powerful hemagglutinin but was only feebly toxic; it emerged from the column as one or more peaks, under different elution conditions. By rechromatography and immunological tests, the toxicity of the second fraction was shown to be due to contamination with traces of the toxic fraction. By several criteria, the toxic and hemagglutinating components appeared to be at least two different proteins.Most of the information obtainable from the ultracentrifugal analysis of these substances was also obtained by gel filtration on a Sephadex G-200 column and with much smaller protein concentrations. At physiological pH, the toxic fraction had a molecular weight of 150,000 and a Stokes radius of 48 A; these dimensions were in agreement with those establishedin vivoby other investigators. The hemagglutinin appeared to exist in three forms of aggregation with molecular weights of 290,000, 500,000, and 900,000.