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首页> 外文期刊>Biochemistry >Complete Subunit Structure of the Clostridium botulinum Type D Toxin Complex via Intermediate Assembly with Nontoxic Components.
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Complete Subunit Structure of the Clostridium botulinum Type D Toxin Complex via Intermediate Assembly with Nontoxic Components.

机译：通过中间组件与无毒组分的中间组件完全胞菌肉毒杆菌型D毒素复合物的完全亚基结构。

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摘要

Clostridium botulinum serotype D strains usually produce two types of stable toxin complex (TC), namely, the 300 kDa M (M-TC) and the 660 kDa L (L-TC) toxin complexes. We previously proposed assembly pathways for both TCs [Kouguchi, H., et al. (2002) J. Biol. Chem. 277, 2650-2656]: M-TC is composed by association of neurotoxin (NT) and nontoxic nonhemagglutinin (NTNHA); conjugation of M-TC with three auxiliary types of hemagglutinin subcomponents (HA-33, HA-17, and HA-70) leads to the formation of L-TC. In this study, we found three TC species, 410, 540, and 610 kDa TC species, in the culture supernatant of type D strain 4947. The 540 and 610 kDa TC species displayed banding patterns on SDS-PAGE similar to that of L-TC but with less staining intensity of the HA-33 and HA-17 bands than those of L-TC, indicating that these are intermediate species in the pathway to L-TC assembly. In contrast, the 410 kDa TC species consisted of M-TC and two molecules of HA-70. All of the TC species, except L-TC, demonstrated no hemagglutination activity. When the intermediate TC species were mixed with an isolated HA-33/17 complex, every TC species converted to 650 kDa L-TC with full hemagglutination activity and had the same molecular composition of L-TC. On the basis of titration analysis with the HA-33/17 complex, the stoichiometry of the HA-33/17 complex molecules in the L-TC, 610 kDa, and 540 kDa TC species was estimated as 4, 3, and 2, respectively. In conclusion, the complete subunit composition of mature L-TC is deduced to be a dodecamer assembled by a single NT, a single NTNHA, two HA-70, four HA-33, and four HA-17 molecules.
机译：Clostridium Botulinum血清型D株通常产生两种类型的稳定毒素复合物（Tc），即300kDa m（M-Tc）和660kDa L（L-Tc）毒素复合物。我们之前提出了TCS的组装途径[Kouguchi，H.等人。 （2002）J. Biol。化学。 277,2650-2656]：M-TC由神经毒素（NT）和无毒性非血凝素（NTNHA）的关联组成; M-TC与三种辅助类型的血凝素子组件（HA-33，HA-17和HA-70）的缀合导致L-Tc的形成。在这项研究中，我们发现了三种TC物种，410,540和610 kDa Tc种类，在D型菌株4947的培养上清液中。540和610 KDA TC物种在类似于L-的SDS-PAGE上显示了绷带图案HA-33和HA-17带的染色强度较少，表明这些是L-TC组件的途径中的中间物质。相反，410kDa Tc物种由M-Tc和两个HA-70分子组成。除了L-TC之外，所有TC种类都证明了没有血凝活性。当中间体TC物种与分离的HA-33/17复合物混合时，每种TC物种转化为650kDa L-Tc，具有全血凝活性，具有相同的L-Tc分子组合物。在用HA-33/17复合物的滴定分析的基础上，L-TC，610kDA和540kDA TC物种中HA-33/17复合物分子的化学计量估计为4,3和2，分别。总之，推导出成熟L-Tc的完全亚基组合物是由单个NT，单个NTNHA，两个HA-70，四HA-33和四个HA-17分子组装的DoDecamer。

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