Characterization of haemagglutinin activity of Clostridium botulinum type C and D 16S toxins, and one subcomponent of haemagglutinin (HA1).
Characterization of haemagglutinin activity of Clostridium botulinum type C and D 16S toxins, and one subcomponent of haemagglutinin (HA1).
复制标题
C 型和 D 型肉毒梭菌 16S 毒素以及血凝素 (HA1) 的一种亚成分的血凝素活性表征。
DOI：
10.1099/00221287-145-9-2533
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发表时间：
1999
期刊：
影响因子：
1.5
通讯作者：
K. Oguma
中科院分区：
文献类型：
--
作者：
K. Inoue;Y. Fujinaga;K. Honke;K. Yokota;T. Ikeda;T. Ohyama;K. Takeshi;T. Watanabe;K. Inoue;K. Oguma
用一种新建立的方法从D型肉毒梭菌培养物中分离纯化了16 S毒素和血凝素（HA）的一个亚组分HA_1，并对它们的HA活性以及纯化的C型16 S毒素的HA活性进行了表征。SDS-PAGE分析表明，游离HA 1形成分子量约为200 kDa的聚合物。C型和D型16 S毒素以相同的方式凝集人红细胞。他们的HA滴度显着降低，通过采用红细胞，已预先处理过的神经氨酸酶，木瓜蛋白酶或蛋白酶K，并通过添加N-乙酰神经氨酸的反应混合物被抑制。在与糖脂如SPG（NeuAc α 2 -3Gal β 1 -4GlcNAc β 1 -3Gal β 1 - 4Glc β 1-Cer）和GM 3（NeuAc α 2 -3Gal β 1 -4Glc β 1-Cer）以及糖蛋白如从红细胞制备的血型糖蛋白A和/或B的直接结合试验中，两种毒素均与唾液酸糖脂和唾液酸糖蛋白结合，但既不与中性糖脂也不与去唾液酸糖蛋白结合。根据这些结果，得出结论，C型和D型165毒素通过N-乙酰神经氨酸与红细胞结合。HA 1没有血凝活性，虽然它结合唾液酸糖脂。因此，我们推测，结合糖蛋白，而不是糖脂可能是重要的C型和D 16 S毒素引起血凝。
The 16S toxin and one subcomponent of haemagglutinin (HA), designated HA1, were purified from a type D culture of Clostridium botulinum by a newly established procedure, and their HA activities as well as that of purified type C 16S toxin were characterized. SDS-PAGE analysis indicated that the free HA1 forms a polymer with a molecular mass of approximately 200 kDa. Type C and D 16S toxins agglutinated human erythrocytes in the same manner. Their HA titres were dramatically reduced by employing erythrocytes that had been previously treated with neuraminidase, papain or proteinase K, and were inhibited by the addition of N-acetylneuraminic acid to the reaction mixtures. In a direct-binding test to glycolipids such as SPG (NeuAc alpha2-3Gal beta1-4GlcNAc beta1-3Gal beta1-4Glc beta1-Cer) and GM3 (NeuAc alpha2-3Gal beta1-4Glc beta1-Cer), and glycoproteins such as glycophorin A and/or B prepared from the erythrocytes, both toxins bound to sialylglycolipids and sialoglycoproteins, but bound to neither neutral glycolipids nor asialoglycoproteins. On the basis of these results, it was concluded that type C and D 165 toxins bind to erythrocytes through N-acetylneuraminic acid. HA1 showed no haemagglutination activity, although it did bind to sialylglycolipids. We therefore speculate that binding to glycoproteins rather than to glycolipids may be important in causing haemagglutination by type C and D 16S toxins.