BLOCKAGE OF TOXIN-CELL INTERACTIONS BY OLIGOSACCHARIDES

我们的目标是设计一种开发抑制剂的通用方案。 细菌毒素与其靶细胞表面的结合 神经鞘糖脂受体。如果毒素与细胞的相互作用被阻断，它 应防止由毒素引起的有害影响 有约束力的。肉毒梭菌神经毒素的结合亲和力 (E型)和破伤风杆菌对其可能的神经节苷脂受体的作用 将被确定是为了确定G1b的哪个神经节苷脂 系列是每种语言的首选接受者。红豆杉的低聚糖部分 破伤风和肉毒杆菌毒素加上GM1的受体 (霍乱弧菌肠道毒素受体)将被分离，使用 本实验室开发的简单、直接的程序，并用于 在载体连接的寡糖和 结合位点定向的不可逆抑制剂。综合方案 建议合成的载体连接的寡糖应为 导致形成簇状低聚糖，可能类似于 当毒素与其细胞表面相互作用时，被毒素“看到”的那些 受体。将对载体连接的低聚糖进行测试 抑制毒素与细胞相互作用的能力。最后，构象。 低聚糖将用核磁共振测定。光谱学和 用于开发毒素-受体相互作用的模型。 这一信息应为制定 位置特异性低聚糖抑制剂，毒素将对其具有 一种高亲和力的毒素-寡糖相互作用 基本上是不可逆转的。

Our aim is to devise a general protocol for the development of inhibitors of the binding of bacterial toxins to their target cell surface glycosphingolipid receptors. If the toxin-cell interaction is blocked, it should prevent the resultant deleterious effects induced by the toxin binding. The binding affinity of the neurotoxins of Clostridium botulinum (type E) and Clostridium tetani to their putative ganglioside receptors will be determined in order to ascertain which ganglioside of the G1b series is the preferred acceptor for each. The oligosaccharide portions of the receptors for the tetanus and botulinum toxin plus that of GM1 (receptor for the enterotoxin of Vibrio cholera) will be isolated, using a simple and straightforward procedure developed in this laboratory, and used in the development of carrier-linked oligosaccharides and binding-site-directed irreversible inhibitors. The synthetic scheme proposed for the synthesis of the carrier-linked oligosaccharides should result in the formation of clustered oligosaccharides possibly resembling those "seen" by the toxin when it interacts with its cell surface receptor. The carrier-linked oligosaccharides will be tested for their ability to inhibit toxin-cell interactions. Finally, the conformation of the oligosaccharides will be determined using n.m.r. spectroscopy and utilized in the development of models of the toxin-receptor interactions. This information should provide the basis for the development of site-specific oligosaccharide inhibitors for which the toxin will have such a high affinity that the toxin-oligosaccharide interaction will be essentially irreversible.