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Purification and Identification of a Novel Protein with Antitumor Activity from Peganum harmala Seeds
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- Author:
- No author available
- Journal Title:
- Natural Product Research and Development
- Issue:
- 8
- DOI:
- 10.3969/j.issn.1001-6880.2012.08.003
- Key Word:
- 骆驼蓬;脂转移蛋白;分离纯化;抗肿瘤;诱导凋亡
Abstract： 骆驼蓬种子经浸提、硫酸铵沉淀、CM阳离子交换层析和Superdex 75凝胶过滤层析分离纯化得到一种具有抗肿瘤细胞增殖活性的蛋白(命名为PhLTP),经Tricine-SDS-PAGE检测为单一蛋白条带,高效液相色谱检测其表观分子量为14.8 kDa左右,表明PhLTP是由两条相同的亚基组成的蛋白.采用Edman降解法对该纯化蛋白的N-末端进行氨基酸测序,其N-末端序列与其他植物非特异性脂转移蛋白相似.对PhLTP抗肿瘤活性进行研究,结果表明其对HeLa、Eca-109、MGC-9和BEL-7404细胞都有增殖抑制活性,其中对HeLa细胞增殖的抑制作用较好,并具有浓度和时间依赖性,其IC50为45 μg/mL.通过Hoechst33258染色观察细胞形态,发现PhLTP能诱导HeLa细胞发生凋亡.
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