肿瘤靶向FoxM1-DBDp蛋白的噬菌体随机肽库筛选与分子模拟
Selection and molecular simulation of phage random peptide library targeting tumor associated FoxM1-DBDp protein
作者：付鹏德;邬怡然;王思怡;李良建;郭红萍;崔健;茆灿泉;
Author：
收稿日期： 年卷（期）页码：2019,34(06):-559-563
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：FoxM1-DBDp;噬菌体随机肽库;生物淘选;多肽;分子对接;反筛测定;细胞生长抑制率;肿瘤
Key words：
基金项目：国家自然科学基金资助项目(批准号:81872789);;
成都市重点研发支撑计划(编号:2018-YF05-00004-SN)
中文摘要
目的从噬菌体随机十二肽库中筛选和获得分子靶向FoxM1-DBDp重组蛋白的高亲和力和肿瘤细胞毒性作用的多肽小分子。方法通过IPTG诱导、原核细菌体外重组表达和Ni-NTA亲和纯化,获得FoxM1-DBDp结合结构域蛋白;以其为靶标对噬菌体随机十二肽库进行四轮生物淘选,获取噬菌斑单克隆,反筛测定阳性克隆与靶蛋白亲和力;采用Discovery Studio 3.0 (DS 3.0)分子模拟对接并用CCK-8测定高亲和力多肽对肿瘤细胞的抑制作用。结果经过四轮生物淘选,高亲和力噬菌体明显得到富集;成功测序了15条序列。反筛测定结果表明:多肽P15具有较高的亲和力;DS分子模拟对接发现该序列与FoxM1-DBD具有最低的结合自由能。CCK-8测定结果显示:100μg·mL~(-1)处理浓度下,P15多肽对肝癌HepG2细胞的抑制率达75.52%。结论通过噬菌体随机十二肽库筛选,成功筛选获得了多条分子靶向FoxM1-DBDp的高亲和力多肽序列,为分子靶向肿瘤密切相关FoxM1的多肽先导药物的研发提供了重要的前期理论基础。
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