Cappuccinelli P, Hames B D
Biochem J. 1978 Mar 1;169(3):499-504. doi: 10.1042/bj1690499.
A colchicine-binding component was detected in vegetative amoebae of Dictyostelium discoideum by using a Millipore-filter assay. The colchicine-binding activity is temperature-and time-dependent, maximum binding occurring at 22-35 degrees C after 60 min incubation. Further increases in temperature are without effect on the extent of binding, but bound colchicine is released with increased time of incubation. Furthermore, colchicine-binding activity itself decreased in the high-speed supernatant from D. discoideum, with half the activity being lost in approx. 2.5h. Several lines of evidence, including the saturation kinetics of colchicine binding, enhancement of colchicine binding by tartrate, insensitivity to lumicolchicine, precipitation of the binding protein by vinblastine and behaviour of the binding protein on DEAE-cellulose and Sephadex resins, suggest that the colchicine-binding protein may be tubulin.
通过使用微孔滤膜分析法，在盘基网柄菌的营养性变形虫中检测到一种秋水仙碱结合成分。秋水仙碱结合活性具有温度和时间依赖性，孵育60分钟后，在22-35摄氏度时结合量达到最大。温度进一步升高对结合程度没有影响，但随着孵育时间延长，结合的秋水仙碱会释放出来。此外，盘基网柄菌高速上清液中的秋水仙碱结合活性本身会降低，约2.5小时内活性丧失一半。包括秋水仙碱结合的饱和动力学、酒石酸盐增强秋水仙碱结合、对光秋水仙碱不敏感、长春碱沉淀结合蛋白以及结合蛋白在二乙氨基乙基纤维素和葡聚糖凝胶树脂上的行为等多条证据表明，秋水仙碱结合蛋白可能是微管蛋白。
