Ramírez G A, Fletcher P L, Possani L D
Department of Biochemistry, UNAM, Cuernavaca, Mexico.
Toxicon. 1990;28(3):285-97. doi: 10.1016/0041-0101(90)90064-e.
The venom from Crotalus molossus nigrescens contains many activities including: hyde powder azure proteinase; N-benzoyl-arginine-ethyl-ester hydrolase; phospholipase; phosphodiesterase; desoxyribonuclease; fibrinogen coagulase; collagenase, fibrinolytic activity, and hemorrhagic factors. The venom, assayed with amounts of venom up to 50 micrograms protein per assay, does not contain acetylcholinesterase, phosphatase, amylase, ribonuclease, tyrosyl-ester hydrolase or hyaluronidase activities. The venom is lethal to mice with an i.p. LD50 of 2.35 mg/kg mouse. Fractionation of soluble venom by Sephadex G-75 separates at least five families of components. Fractions I-III contains all the enzymes, and fraction V have six small peptides. Further separation of fractions II-III on diethyl-amino-ethyl-cellulose columns at pH 8.0 and 8.3 gave pure proteinase E with a mol. wt of 21,390 and the following N-terminal amino acid sequence; Phe-Ala-Lys-Arg-Tyr-Val-Glx-Leu-Val-Ile-Val-Ala. A thrombin-like enzyme with a mol. wt of 75,000 was also purified from this venom by means of affinity and ion exchange chromatographies.
黑尾拟眼镜蛇的毒液含有多种活性成分，包括：透明质酸酶；N-苯甲酰-精氨酸乙酯水解酶；磷脂酶；磷酸二酯酶；脱氧核糖核酸酶；纤维蛋白原凝固酶；胶原酶、纤维蛋白溶解活性和出血因子。用每次测定高达50微克蛋白质的毒液量进行检测时，该毒液不含乙酰胆碱酯酶、磷酸酶、淀粉酶、核糖核酸酶、酪氨酰酯水解酶或透明质酸酶活性。该毒液对小鼠具有致死性，腹腔注射的半数致死量为2.35毫克/千克小鼠。用葡聚糖凝胶G-75对可溶性毒液进行分级分离，可分离出至少五个组分家族。组分I-III包含所有酶，组分V有六个小肽。在pH 8.0和8.3条件下，在二乙氨基乙基纤维素柱上对组分II-III进行进一步分离，得到分子量为21,390的纯蛋白酶E，其N端氨基酸序列如下：苯丙氨酸-丙氨酸-赖氨酸-精氨酸-酪氨酸-缬氨酸-谷氨酰胺-亮氨酸-缬氨酸-异亮氨酸-缬氨酸-丙氨酸。还通过亲和色谱和离子交换色谱从这种毒液中纯化出一种分子量为75,000的凝血酶样酶。
