Identification of the interaction region between hemagglutinin components of the botulinum toxin complex
Identification of the interaction region between hemagglutinin components of the botulinum toxin complex
复制标题
肉毒杆菌毒素复合物的血凝素成分之间相互作用区域的鉴定
DOI：
10.1016/j.ijbiomac.2014.01.052
复制
发表时间：
2014
影响因子：
8.2
通讯作者：
Sagane Y
中科院分区：
文献类型：
--
作者：
Suzuki T;Miyashita S;Hayashi S;Miyata K;Inui K;Kondo Y;Miyazaki S;Ohyama T;Niwa K;Watanabe T;Sagane Y
肉毒梭菌（Clostridium botulinum）产生的大毒素复合物（L-TC）是由M-TC与HA-33/HA-17三聚体偶联形成的，由两个HA-33蛋白和一个HA-17蛋白组成。这种结合由HA-70介导，HA-70与HA-17相互作用。目前的研究旨在鉴定HA-70分子中粘附于HA-33/HA-17复合物的区域。通过SDS-PAGE解析来自HA-70的有限蛋白水解的产物，并转移到PVDF膜上，在那里它们在远蛋白质印迹中用HA-33/HA-17探测。在HA-70片段中，HA-33/HA-17与至少含有HA-70分子的C-末端一半的那些片段结合，但不与携带N-末端一半的那些片段结合。另外的对接模拟分析表明，HA-70区域Gln 420-Tyr 575负责与HA-17结合，这与远蛋白质印迹数据一致。这里的研究结果揭示了有关肉毒杆菌毒素复合物中功能性HA子复合物的三维结构的更多细节。
The large toxin complex (L-TC) produced byClostridium botulinumis formed from the M-TC (BoNT/NTNHA complex) by conjugation of M-TC with HA-33/HA-17 trimer consists of two HA-33 proteins and a single HA-17 protein. This association is mediated by HA-70, which interacts with HA-17. The current study aims to identify the regions of the HA-70 molecule that adhere to the HA-33/HA-17 complex. Products from limited proteolysis of HA-70 were resolved by SDS-PAGE and transferred onto PVDF membranes, where they were probed with HA-33/HA-17 in a far-western blot. Among the HA-70 fragments, HA-33/HA-17 bound to those containing at least the C-terminal half of the HA-70 molecule, but not those carrying the N-terminal half. Additional docking simulation analysis indicated that the HA-70 region Gln420-Tyr575 is responsible for binding to HA-17, which is consistent with the far-western blot data. The findings here reveal additional details concerning the three-dimensional structure of the functional HA sub-complex in the botulinum toxin complex.