Characterization of toxin complex produced by a unique strain of Clostridium botulinum serotype D 4947
Characterization of toxin complex produced by a unique strain of Clostridium botulinum serotype D 4947
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DOI：
10.1023/b:jopc.0000039551.42676.4c
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发表时间：
2004-08-01
期刊：
影响因子：
3
通讯作者：
Ohyama, T
中科院分区：
文献类型：
--
作者：
Hasegawa, K;Watanabe, T;Ohyama, T
一种独特的肉毒杆菌菌株，血清型D 4947 (D-4947)，产生大量650 kDa的毒素复合物（L-TC）和少量280 kDa的M-TC， 540 kDa的TC和610 kDa的TC。这些复合物仅由非缺口成分组成，包括神经毒素（NT）、无毒的非血凝素（NTNHA）和血凝素亚成分（HA-70、HA-33和HA-17）。与所有血清型菌株的其他NT不同，除M-TC外，在色谱过程中，D-4947 NT与L-TC的分离需要pH 8.8左右的高碱性条件。分离的NT和NTNHA/HAs复合物可以重组成L-TC，在毒性、血凝活性和凝胶过滤特性方面与母体L-TC没有区别。根据HA-33/17分子数的不同，NT和NTNHA/HAs的等电点接近。我们建立了一种新的方法来分离独特的D-4947 NT，这将为肉毒毒素的结构提供有价值的信息。
A unique strain of Clostridium botulinum, serotype D 4947 (D-4947), produces a considerable amount of a 650 kDa toxin complex (L-TC) and a small amount of a 280 kDa M-TC, a 540 kDa TC, and a 610 kDa TC. The complexes are composed of only un-nicked components, including neurotoxin (NT), nontoxic nonhemagglutinin (NTNHA) and hemagglutinin subcomponents (HA-70, HA-33 and HA-17). Unlike other NTs from all serotype strains, separation of D-4947 NT from L-TC, except for M-TC, during chromatography required highly alkaline conditions around pH 8.8. The separated NT and NTNHA/HAs complex can be reconstituted to L-TC that is indistinguishable from the parent L-TC with respect to toxicity, hemagglutination activity and gel filtration profile. The isoelectric points of NT and NTNHA/HAs were close together depending on the number of HA-33/17 molecules. We have established a new method to separate the unique D-4947 NT from the complex, which will yield valuable information on structure of botulinum toxin.