Aird S D, Kaiser I I
Toxicon. 1985;23(3):361-74. doi: 10.1016/0041-0101(85)90020-0.
Toxins from the venoms of Crotalus durissus terrificus, Crotalus s. scutulatus and Crotalus viridis concolor were compared using gel filtration, ion-exchange chromatography on DEAE-Sephacel and denaturing and non-denaturing polyacrylamide gel electrophoresis. The three heterodimeric native toxins behaved similarly on each of the separation media, except that the C. d. terrificus toxin displayed a pronounced tendency to dissociate on DEAE-Sephacel, even in the absence of urea. In the presence of 6M urea, subunit dissociation was quantitative for all three toxins. Recombination of purified subunits resulted in toxins which eluted from the gel filtration column in identical fashion to native toxins. Non-denaturing polyacrylamide gel electrophoretic patterns of recombined toxins actually showed greater band resolution than did the native toxins. Six hybrid toxins were generated on polyacrylamide gels from cross-combinations of purified subunits, each with different mobilities than the parental toxins. Mobilities of the hybrid toxins depended principally upon the mobilities of the basic subunits. All three purified native toxins showed comparable LD50's in female mice (0.039-0.061 micrograms/g). The C. d. terrificus acidic X C. s. scutulatus basic hybrid toxin showed toxicity identical to that of the C. s. scutulatus recombined toxin. Phospholipase activity is associated with the basic subunit in all three toxins. Intact toxins show a distinctive lag in phospholipase activity which is not seen with purified basic subunits alone. These results indicate that the principal toxins in these three venoms are homologous.
使用凝胶过滤、DEAE - 葡聚糖凝胶离子交换色谱以及变性和非变性聚丙烯酰胺凝胶电泳对杜氏响尾蛇（Crotalus durissus terrificus）、盾形响尾蛇（Crotalus s. scutulatus）和同色响尾蛇（Crotalus viridis concolor）毒液中的毒素进行了比较。这三种异源二聚体天然毒素在每种分离介质上的表现相似，只是杜氏响尾蛇毒素即使在没有尿素的情况下，在DEAE - 葡聚糖凝胶上也有明显的解离倾向。在6M尿素存在下，所有三种毒素的亚基解离都是定量的。纯化亚基的重组产生的毒素从凝胶过滤柱上洗脱的方式与天然毒素相同。重组毒素的非变性聚丙烯酰胺凝胶电泳图谱实际上比天然毒素显示出更高的条带分辨率。在聚丙烯酰胺凝胶上通过纯化亚基的交叉组合产生了六种杂交毒素，每种杂交毒素的迁移率都与亲本毒素不同。杂交毒素的迁移率主要取决于碱性亚基的迁移率。所有三种纯化的天然毒素在雌性小鼠中的半数致死剂量（LD50）相当（0.039 - 0.061微克/克）。杜氏响尾蛇酸性亚基×盾形响尾蛇碱性亚基杂交毒素的毒性与盾形响尾蛇重组毒素相同。所有三种毒素的磷脂酶活性都与碱性亚基相关。完整毒素在磷脂酶活性上表现出独特的滞后现象，而单独的纯化碱性亚基则没有这种现象。这些结果表明这三种毒液中的主要毒素是同源的。
