Jiménez-González A, De Armas-Serra C, Criado-Fornelio A, Casado-Escribano N, Rodríguez-Caabeiro F, Díez J C
Parasitology Laboratory, Faculty of Pharmacy, University of Alcaldá de Henares, Madrid, Spain.
Vet Parasitol. 1991 Jul;39(1-2):89-99. doi: 10.1016/0304-4017(91)90065-4.
Tubulin was estimated to account for 0.3% of the total soluble protein in Trichinella spiralis cytosolic fractions. Tubulin from T. spiralis was partially purified by precipitation with either taxol or vinblastine sulphate. Immunoblotting with alpha- and beta-tubulin monoclonal antibodies revealed the presence of tubulin in T. spiralis partially purified preparations. Electrophoretic mobility of T. spiralis tubulin in sodium dodecyl sulphate-polyacrylamide gels was very similar to that shown by pig brain tubulin. Further studies with colchicine binding assays indicated that T. spiralis tubulin has binding features similar to that of tubulin from other nematodes: colchicine association constant = 8.1 x 10(-4) M and competitive inhibition of colchicine binding by podophyllotoxin, with an inhibition constant of 1.3 x 10(-6) M. Finally, inhibition of colchicine binding by several benzimidazoles (mebendazole, fenbendazole, oxibendazole and albendazole) was investigated. All the benzimidazoles inhibited colchicine binding in a competitive manner, with inhibition constant values ranging from 1.4 x 10(-7) M (mebendazole) to 3.9 x 10(-6) M (fenbendazole).
据估计，微管蛋白占旋毛虫胞质组分中总可溶性蛋白的0.3%。旋毛虫的微管蛋白通过用紫杉醇或硫酸长春碱沉淀进行部分纯化。用α-和β-微管蛋白单克隆抗体进行免疫印迹分析，结果显示在旋毛虫部分纯化的制剂中存在微管蛋白。在十二烷基硫酸钠-聚丙烯酰胺凝胶中，旋毛虫微管蛋白的电泳迁移率与猪脑微管蛋白的电泳迁移率非常相似。用秋水仙碱结合试验进行的进一步研究表明，旋毛虫微管蛋白具有与其他线虫微管蛋白相似的结合特性：秋水仙碱缔合常数 = 8.1×10⁻⁴ M，鬼臼毒素对秋水仙碱结合具有竞争性抑制作用，抑制常数为1.3×10⁻⁶ M。最后，研究了几种苯并咪唑类药物（甲苯咪唑、芬苯达唑、奥苯达唑和阿苯达唑）对秋水仙碱结合的抑制作用。所有苯并咪唑类药物均以竞争性方式抑制秋水仙碱结合，抑制常数范围从1.4×10⁻⁷ M（甲苯咪唑）到3.9×10⁻⁶ M（芬苯达唑）。
