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用两个循环聚合——解聚的方法,从猪脑分离到部分纯化的微管蛋白,在聚丙烯酰胺凝胶电泳图谱上显示两个主峰带,即。微管蛋白(M:W.58,000)和β微管蛋白(M.W.54,000)。在ATP或GTP存在的MES缓冲液中37℃保温b微管蛋白迅速聚合为微管,A 350呈典型S形聚合曲线。在电子显微镜下显示网状微管结构。抗癌药物筛选实验表明:秋水仙碱明显抑制微管聚合;长春新碱低浓度(0.75-6μM)抑制微管蛋白聚合,高浓度(>6μM)时导致微管蛋白聚集(Aggregation),且在低温下不解聚。三尖杉酯类生物碱对微管蛋白的聚合及解聚未显示明显的影响。
Two cycles of polymerization-depolymerization were used to isolate partially purified tubulin from porcine brain, showing two major bands on a polyacrylamide gel electrophoresis pattern. Tubulin (M: W. 58,000) and β-tubulin (M. W. 54,000). Incubation at 37°C in the presence of ATP or GTP in MES buffer, b-tubulin rapidly polymerized into microtubules, and A 350 showed a typical S-shaped polymerisation curve. The mesh microtubule structure is shown under an electron microscope. Anti-cancer drug screening experiments showed that: colchicine significantly inhibited microtubule polymerization; vincristine low concentration (0.75-6μM) inhibition of tubulin polymerization, high concentrations (> 6μM) lead to tubulin aggregation (Aggregation), and in Do not disaggregate at low temperatures. The harringtonine alkaloids showed no significant effect on the polymerization and depolymerization of tubulin.