IMMUNOLOGICAL CHARACTERIZATION OF PAPAIN-INDUCED FRAGMENTS OF CLOSTRIDIUM-BOTULINUM TYPE-A NEUROTOXIN AND INTERACTION OF THE FRAGMENTS WITH BRAIN SYNAPTOSOMES
IMMUNOLOGICAL CHARACTERIZATION OF PAPAIN-INDUCED FRAGMENTS OF CLOSTRIDIUM-BOTULINUM TYPE-A NEUROTOXIN AND INTERACTION OF THE FRAGMENTS WITH BRAIN SYNAPTOSOMES
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DOI：
10.1128/iai.57.9.2634-2639.1989
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发表时间：
1989-09-01
影响因子：
3.1
通讯作者：
SAKAGUCHI, G
中科院分区：
文献类型：
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作者：
KOZAKI, S;MIKI, A;SAKAGUCHI, G
用木瓜蛋白酶处理A型肉毒梭菌神经毒素后，通过疏水色谱和离子交换色谱与高效液相色谱系统纯化三个片段（Mrs，101，000，45，000和43，000）。用单克隆抗体进行的免疫印迹分析表明，101，000-道尔顿片段由通过二硫键连接在一起的轻链和重链的一部分（H-1片段）组成，另外两个片段与重链的剩余部分（H-2片段）相关。45，000-和43，000-道尔顿片段有效地竞争125 I标记的神经毒素与突触体的结合，而101，000-道尔顿片段没有观察到抑制作用。结果表明，H-2片段与神经膜上的结合位点相互作用。神经毒素的结合受损的突触体与神经氨酸酶的治疗。将神经节苷脂掺入神经氨酸酶处理的突触体中导致结合的恢复。结果表明，神经节苷脂是毒素结合位点的成分之一。
After treatment of Clostridium botulinum type A neurotoxin with papain, three fragments (Mrs, 101,000, 45,000, and 43,000) were purified by hydrophobic and ion-exchange chromatography with a high-performance liquid chromatographic system. Immunoblotting analyses with monoclonal antibodies showed that the 101,000-dalton fragment consisted of the light chain and a part of the heavy chain (H-1 fragment) linked together by a disulfide bond, and the other two fragments were correlated to the remaining portion of the heavy chain (H-2 fragment). The 45,000- and 43,000-dalton fragments effectively competed for binding of the 125I-labeled neurotoxin to synaptosomes, while no inhibition was observed with the 101,000-dalton fragment. The results indicate that the H-2 fragment interacts with the binding site on the neural membrane. The binding of the neurotoxin was impaired by treatment of synaptosomes with neuraminidase. Incorporation of gangliosides into neuraminidase-treated synaptosomes resulted in the restoration of binding. The results suggest that gangliosides are one of the components of the toxin-binding site.