Ivanova L G, Blagoveshchenskiĭ V A, Vinogradova I D, Kolesnikova V A, Ugriumova G A
Biokhimiia. 1983 Sep;48(9):1548-54.
A procedure for isolation of highly purified hemagglutinin from a toxic complex of culture filtrates of Cl. botulinum type A is described. This procedure includes precipitation with (NH4)2SO4, chromatography on Sephadex G-100, G-200 and DEAE-cellulose, specific adsorption on human erythrocytes and affinity chromatography. Using polyacrylamide gel electrophoresis, it was shown that hemagglutinin is a heteropolymeric protein consisting of a monomer (Mr 53 000) and a trimer (Mr 160 000). The monomer is made up of two subunits with Mr 13 000 and one subunit with Mr 27 000 covalently linked by SS-crosslinks. The number and nature of the SS-crosslinks and SH-groups in the protein molecule were determined and a hypothetical structural model of hemagglutinin was proposed. Using immunochemical analysis, it was shown that some (but not all) serological properties of the highly purified protein from Cl. botulinum type A and of its partially purified counterpart are similar to those of hemagglutinin from Cl. botulinum type B.
本文描述了一种从A型肉毒杆菌培养滤液的毒性复合物中分离高纯度血凝素的方法。该方法包括用硫酸铵沉淀、在葡聚糖G - 100、G - 200和二乙氨基乙基纤维素上进行色谱分离、在人红细胞上的特异性吸附以及亲和色谱。通过聚丙烯酰胺凝胶电泳表明，血凝素是一种异聚蛋白，由一个单体（分子量53000）和一个三聚体（分子量160000）组成。单体由两个分子量为13000的亚基和一个分子量为27000的亚基通过二硫键交联共价连接而成。确定了蛋白质分子中二硫键交联和巯基的数量和性质，并提出了血凝素的假设结构模型。通过免疫化学分析表明，A型肉毒杆菌高纯度蛋白及其部分纯化对应物的一些（但不是全部）血清学特性与B型肉毒杆菌血凝素的血清学特性相似。
