蛇毒抗肿瘤蛋白心脏毒素的分离纯化与鉴定

Separation Purification and identification of cardiotoxin in atranticance protein

目的:对蛇毒抗瘤蛋白的活性组分进行分离与鉴定。方法:采用 RP-HPLC 分离蛇毒抗瘤蛋白各组分,收集纯化组分Ⅱ。RP-HPLC 及 SDS-PAGE 测定组分Ⅱ的纯度;MALDI—TOF 质谱仪测定组分Ⅱ的相对分子质量;对组分Ⅱ进行 N-末端氨基酸序列测定。结果:蛇毒抗瘤蛋白组分Ⅱ经 RP-HPLC 和 SDS-PAGE 分析为单一成分,MALDI-TOF 质谱仪测得其相对分子质量为6719.49,其 N-末端氨基酸序列为 L-K-C-N-K-L-V-P-L-F-Y-K-T-C-P。结论:经分离鉴定,蛇毒抗瘤蛋白组分Ⅱ为心脏毒素。

Objective： Isolation, purification and identification of cardiotoxin in atranticance protein. Methods： Fractions of atranticance protein were isolated by RP - HPLC and the fraction Ⅱwas purified. Purity of the fraction Ⅱ was determined by RP - HPLC and SDS - PAGE. The relative molecular mass was determined by MALDI - TOF. N - teminal sequence of the fraction Ⅱ was analyzed. Results： The fraction Ⅱ was verified to be homogeneous by RP - HPLC and SDS - PAGE. The relative molecular mass of the fraction Ⅱ was 6719.49 by MALDI - TOF. The results of N - teminal sequence analysis were L - K - C - N - K - L - V - P - L - F - Y - K - T - C - P. Conclusion：The fraction Ⅱ in atranticance protein is cardiotoxin.