Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms
Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms
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DOI：
10.1128/iai.65.5.1626-1630.1997
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发表时间：
1997-05-01
影响因子：
3.1
通讯作者：
Stevens, RC
中科院分区：
文献类型：
--
作者：
Chen, F;Kuziemko, GM;Stevens, RC
通过使用 44 个单克隆单链可变片段的抗体图谱研究了肉毒杆菌神经毒素 A 血清型的结构域组织，分析对 (i) 肉毒杆菌神经毒素全毒素的各个结构域（结合、易位和催化）、(ii) 肉毒杆菌神经毒素全毒素、(iii) 与无毒部分复合的肉毒杆菌神经毒素全毒素和 (iv) 进行分析。肉毒杆菌神经毒素全毒素和复合物的无毒部分在体外重组。所有 44 种抗体都映射到肉毒杆菌神经毒素的各个结构域。 44 个单链可变片段中的 40 个相对于分离的结构域结合肉毒杆菌神经毒素全毒素，表明当各个结构域处于全毒素形式时覆盖了 4 个表位。只有 20 种抗体在与无毒部分复合时显示出对毒素的阳性反应。所有覆盖的表位都映射到肉毒杆菌神经毒素的结合域，这表明结合域与复合物中的无毒部分直接接触。基于映射到肉毒杆菌神经毒素全毒素和整个复合物的不同域的抗体，提出了肉毒杆菌神经毒素复合物的模型。
The domain organization of the botulinum neurotoxin serotype A was studied by using antibody mapping of 44 monoclonal single-chain variable fragments, The analysis was carried out on (i) the individual domains of botulinum neurotoxin holotoxin (binding, translocation, and catalytic), (ii) botulinum neurotoxin holotoxin, (iii) the botulinum neurotoxin holotoxin in complex with the nontoxic portion, and (iv) botulinum neurotoxin holotoxin and nontoxic portion of the complex recombined in vitro. All 44 antibodies mapped to individual domains of botulinum neurotoxin. Forty of the 44 single-chain variable fragments bound the botulinum neurotoxin holotoxin relative to the isolated domains, suggesting that 4 epitopes are covered when the individual domains are in the holotoxin form. Only 20 of the antibodies showed a positive reaction to the toxin while in complex with the nontoxic portion. All of the covered epitopes were mapped to the binding domain of botulinum neurotoxin, which suggested that the binding domain is in direct contact with the nontoxic portion in the complex, Based on the antibody mapping to the different domains of the botulinum neurotoxin holotoxin and the entire complex, a model of the botulinum neurotoxin complex is proposed.