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5. 新型直接凝血酶抑制剂短肽(DTIP)重要氨基酸研究

新型直接凝血酶抑制剂短肽(DTIP)重要氨基酸研究

来源 :中国生物化学与分子生物学会第十二届会员代表大会暨2018年全国学术会议 | 被引量 : 0次 | 上传用户：zhangyong131420

【摘 要】

：

目的 研究影响 DTIP 稳定性及与凝血酶相互作用的氨基酸并进行结构改造。方法 通过 discovery studio 同源建模构建 DTIP 三维结构模型，与凝血酶分子对接，通过丙氨酸扫描筛选突变能不稳定的氨基酸；其次通过单氨基酸突变，获得 DTIP 突变体；用凝血酶滴定法测定各突变体抗凝血酶活性，利用小鼠模型测定部分凝血酶活酶时间、凝血酶时间、凝血酶原时间，剪尾实验测定出血时间，通过生物膜层干

【作 者】

：

李亚然 于敏 莫炜 

【机 构】

：

复旦大学代谢与分子医学教育部重点实验室 上海 200032;复旦大学基础医学院生 物化学与分子生物学系 上海 200032

【出 处】

：

中国生物化学与分子生物学会第十二届会员代表大会暨2018年全国学术会议

【发表日期】

：

2018年10期

【关键词】

：

DTIP 丙氨酸扫描 凝血酶 血栓 

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　　目的 研究影响 DTIP 稳定性及与凝血酶相互作用的氨基酸并进行结构改造。方法 通过 discovery studio 同源建模构建 DTIP 三维结构模型，与凝血酶分子对接，通过丙氨酸扫描筛选突变能不稳定的氨基酸；其次通过单氨基酸突变，获得 DTIP 突变体；用凝血酶滴定法测定各突变体抗凝血酶活性，利用小鼠模型测定部分凝血酶活酶时间、凝血酶时间、凝血酶原时间，剪尾实验测定出血时间，通过生物膜层干涉实验测定各突变蛋白与凝血酶相互作用的亲和力，血栓弹力实验检测其对全血的影响。

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