蛇毒抗肿瘤蛋白化学成分的研究

Study on the Chemical Component in Atranticance Protein

对蛇毒抗肿瘤蛋白的化学成分进行研究。采用RP-HPLC分离蛇毒抗肿瘤蛋白各组分,收集纯化组分Ⅳ。非还原型SDS-PAGE和RP-HPLC鉴定组分Ⅳ的纯度;还原型SDS-PAGE与MALDI-TOF质谱仪测定组分Ⅳ的相对分子质量;测定组分ⅣN-末端氨基酸序列并进行Western blot鉴定;MTT法检测组分Ⅳ对体外培养的K562细胞的生长抑制作用。结果表明蛇毒抗肿瘤蛋白组分Ⅳ经SDS-PAGE和RP-HPLC分析为单一成分,MALDI-TOF质谱仪测得其相对分子质量为6714.22,其N-末端前5个氨基酸序列为L-K-C-N-K。经Western blot鉴定蛇毒抗肿瘤蛋白组分Ⅳ为细胞毒素,其对K562细胞的抑制作用呈明显的剂量依赖关系。

Order to Study on the chemical component in Atranticance Protein （AP）, fractions of AP were isolated by RP-HPLC and the AP-fractionⅣwas purified. Purity of the AP- fractionⅣwas determined by SDS-PAGE and RP-HPLC, the molecular weight was determined by MALDI-TOF, then Nteminal sequences were analyzed. The AP- fractionⅣwas determined by Western blot, the inhibition of the AP- fraction]Yon K562 was examined by MTT assay. The results showed that the AP-fractionⅣwas homogeneous, the molecular weight was 6714.22, N-teminal sequences was L-K-C-N-K. The AP- fractionⅣ was a cytotoxin by Western blot. The inhibitory effect of the AP- fractionⅣ on K562 showed a definite dosage-dependent relationship.