Analysis of receptor recognition site on botulinum neurotoxin molecule and its interaction with the receptor.

肉毒神经毒素分子受体识别位点及其与受体相互作用的分析。

• 批准号: 11670268
• 负责人: KOZAKI Shunji
• 金额: $ 2.24万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670268/
• 关键词: Clostridium botulinum; neurotoxin; receptor; mutant

We examined properties of the neurotoxin (111/NT) produced by the organism which had been associated with the first type B infant botulism and compared with the nerutoxin (Okra/NT) of the authentic strain. The data indicate that the toxicity of 111/NT was lower than that of Okra/NT, being due to the receptor binding capability. In fact the neucleotide sequence analysis suggest that the receptor binding domains between the two toxins share different amino acid residues and about 70% of these residues expect to be exposed to the surface on the toxin molecule. We are now in progress to examine the the receptor binding abilities of various mutants of the toxin binding domain in order to clarify the recognition site.Synaptotagmin II is the protein receptor for the neurotoxin and has the toxin binding site on the amine-terminal region involving 60 amino acid residues. Overall, 40 site-directed and deleted mutants were prepared to examine their toxin binding activity. The data showed that six amino acid residues are related with the toxin binding. Such critical amino acid residues are located in the 41 to 60 residues from the amino termius, indicating that the 20 residues function as the toxin recognition site.

我们检查了与第一种 B 型婴儿肉毒杆菌中毒相关的生物体产生的神经毒素 (111/NT) 的特性，并与真实菌株的神经毒素 (Okra/NT) 进行了比较。数据表明，由于受体结合能力，111/NT 的毒性低于 Okra/NT。事实上，核苷酸序列分析表明，两种毒素之间的受体结合结构域共享不同的氨基酸残基，并且这些残基中的约 70% 预计会暴露在毒素分子的表面。我们现在正在检查毒素结合结构域的各种突变体的受体结合能力，以澄清识别位点。突触结合蛋白 II 是神经毒素的蛋白质受体，在胺末端区域具有毒素结合位点，涉及60个氨基酸残基。总体而言，制备了 40 个定点突变体和缺失突变体来检查它们的毒素结合活性。数据显示，有六个氨基酸残基与毒素结合有关。这样的关键氨基酸残基位于从氨基端算起的41至60个残基，表明这20个残基充当毒素识别位点。

项目成果

小崎 俊司: "ボツリヌス神経毒素の受容体に関する研究"日本細菌学雑誌. 54. 401-414 (1999)

Shunji Ozaki：“肉毒杆菌神经毒素受体的研究”日本细菌学杂志 54. 401-414 (1999)。

Kamata, Y.: "Difference in hydrophobicity between botulinum type B activated and non-activated neurotoxins under low pH conditions."Toxicon. 38. 1247-1251 (2000)

Kamata, Y.：“低 pH 条件下 B 型肉毒杆菌激活和非激活神经毒素之间的疏水性差异。”Toxicon。

小崎俊司: "ボツリヌス神経毒素の受容体に関する研究"日本細菌学雑誌. 54. 401-414 (1999)

Shunji Ozaki：“肉毒杆菌神经毒素受体的研究”日本细菌学杂志 54. 401-414 (1999)。

Kohda,T.: "Endocytosis of Clostridium botulinum type B neurotoxin into rat brain synaptosomes."J.Vet.Med.Sci.. 62. 1133-1138 (2000)

Kohda,T.：“B 型肉毒杆菌神经毒素内吞到大鼠脑突触体中。”J.Vet.Med.Sci.. 62. 1133-1138 (2000)

Kozaki, S.: "Studies on the receptor for Clostridium botulinum neurotoxin (In Japanese)"Nihon Saikingaku Zatsushi. 54. 401-414 (1999)

Kozaki, S.：“肉毒杆菌神经毒素受体的研究（日语）”Nihon Saikingaku Zatsushi。