The use of monoclonal antibodies to analyze the structure of Clostridium botulinum type E derivative toxin
The use of monoclonal antibodies to analyze the structure of Clostridium botulinum type E derivative toxin
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利用单克隆抗体分析E型肉毒梭菌衍生毒素的结构
DOI：
10.1128/iai.52.3.786-791.1986
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发表时间：
1986
影响因子：
3.1
通讯作者：
G. Sakaguchi
中科院分区：
文献类型：
--
作者：
S. Kozaki;Y. Kamata;T. Nagai;J. Ogasawara;G. Sakaguchi
制备了6株抗E型肉毒梭菌衍生毒素的单克隆抗体。与重链结合的五个中的三个中和衍生毒素;与轻链结合的另一个没有。用单克隆抗体进行的免疫印迹分析表明，通过胰蛋白酶消化获得的片段由通过二硫键连接在一起的轻链和部分重链（H-1片段）组成，并且E型和F型衍生毒素之间共有的抗原决定簇位于重链和轻链上。糜蛋白酶处理诱导的片段，像胰蛋白酶片段，结合到四个单克隆抗体。温和的胰蛋白酶处理和还原导致胰凝乳蛋白酶片段分离成两个较小的片段，对应于轻链和H-1片段。这些结果表明H-1片段含有重链的氨基末端部分。中和毒素的单克隆抗体，并可能识别的羧基末端部分（H-2片段）的重链上的表位有效地竞争结合125 I标记的衍生毒素突触体。中和毒素并识别H-1片段上的表位的两种单克隆抗体中，一种部分抑制结合，而另一种不抑制结合。这表明125 I标记的衍生毒素的结合主要取决于重链的羧基末端区域，并且干扰结合不是毒素中和的唯一手段。
Six monoclonal antibodies against Clostridium botulinum type E derivative toxin were prepared. Three of the five binding to the heavy chain neutralized the derivative toxin; the other one binding to the light chain did not. Immunoblotting analysis with the monoclonal antibodies showed that the fragment obtained by tryptic digestion consisted of the light chain and part of the heavy chain (H-1 fragment) linked together by a disulfide bond(s) and that the antigenic determinants common between type E and F derivative toxins were located on both the heavy and light chains. The fragment induced by chymotrypsin treatment, like the tryptic fragment, bound to four monoclonal antibodies. The mild tryptic treatment and reduction resulted in separation of the chymotryptic fragment into two smaller fragments corresponding to the light chain and H-1 fragment. These results indicate that H-1 fragment contains the amino-terminal portion of the heavy chain. The monoclonal antibody neutralizing the toxin and probably recognizing the epitope on the carboxyl-terminal portion (H-2 fragment) of the heavy chain effectively competed for binding of 125I-labeled derivative toxin to synaptosomes. Of the two monoclonal antibodies neutralizing the toxin and recognizing the epitopes on H-1 fragment, one partially inhibited binding, but the other did not. This suggests that the binding of 125I-labeled derivative toxin depends mainly on the carboxyl-terminal region of the heavy chain and that interference with binding is not the only means of toxin neutralization.
E型肉毒杆菌神经毒素重链和轻链的部分氨基酸序列。
DOI：
10.1016/s0006-291x(85)80009-7
发表时间：
1985
影响因子：
3.1
作者：
Sathyamoorthy,V;DasGupta,BR
通讯作者：
DasGupta,BR
肉毒杆菌毒素的起源、结构和药理活性。
影响因子：
21.1
作者：
Simpson,LL
通讯作者：
Simpson,LL