Hasegawa Kimiko, Watanabe Toshihiro, Sato Hiroaki, Sagane Yoshimasa, Mutoh Shingo, Suzuki Tomonori, Yamano Akihito, Kouguchi Hirokazu, Takeshi Kouichi, Kamaguchi Arihide, Fujinaga Yukako, Oguma Keiji, Ohyama Tohru
Department of Food Science and Technology, Faculty of Bioindustry, Tokyo University of Agriculture, Abashiri 099-2493, Japan.
Protein J. 2004 Aug;23(6):371-8. doi: 10.1023/b:jopc.0000039551.42676.4c.
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.
一种独特的肉毒杆菌菌株，D型4947株（D - 4947），能产生大量的650 kDa毒素复合物（L - TC）以及少量的280 kDa M - TC、540 kDa TC和610 kDa TC。这些复合物仅由未切割的成分组成，包括神经毒素（NT）、无毒非血凝素（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NTs不同，在色谱分析过程中，从L - TC（M - TC除外）中分离D - 4947 NT需要pH值约为8.8的高碱性条件。分离出的NT和NTNHA/HAs复合物可以重新组装成L - TC，其毒性、血凝活性和凝胶过滤图谱与亲本L - TC无法区分。NT和NTNHA/HAs的等电点彼此接近，这取决于HA - 33/17分子的数量。我们已经建立了一种从复合物中分离独特的D - 4947 NT的新方法，这将为肉毒杆菌毒素的结构提供有价值的信息。
