胱硫醚β合酶天然产物抑制剂的高通量筛选

发布时间：2018-12-17 00:55

【摘要】：[目的]构建人源胱硫醚β合酶CBS△516-525体外高通量筛选模型,在基础酶活水平,筛选天然产物库,发现新型、高效且特异的CBS天然产物抑制剂。[方法]在192串联双孔板内,构建纯酶CBS△516-525的高通量筛选模型,以200!mol/L初筛5 206个天然产物,经50!mol/L和5!mol/L复测验证后,选择抑制效果好的新型抑制剂进行剂量依赖性测试并检测其对同源酶胱硫醚γ裂解酶(CSE)的抑制效果。[结果]原核表达纯化出具有生物活性的CBS△516-525酶,经高通量筛选发现2个IC50约1!mol/L的CBS天然产物抑制剂,且对同源酶CSE均有180倍选择性。[结论]在基础酶活条件下,高通量筛选发现了选择性大于180倍的CBS高效天然产物抑制剂,这可为CBS的分子机制和功能研究提供特异的分子工具。
[Abstract]:[objective] to construct a high throughput screening model of human cystathionine 尾 synthase CBS 516-525 in vitro, and to screen the natural product library at the basic enzyme activity level, and to find a novel, efficient and specific inhibitor of CBS natural product. [methods] A high throughput screening model of pure enzyme CBS 516-525 was constructed in a 192 tandem double-hole plate. 5206 natural products were screened with 200g mol / L, and the results were verified by repeated measurements of 50 mol / L and 5 mol / L, respectively. A new inhibitor with good inhibitory effect was selected for dose-dependent test and its inhibitory effect on homologous cystathithione 纬 lyase (CSE) was tested. [results] the CBS 516-525 enzyme with biological activity was purified from prokaryotic expression. Two CBS natural product inhibitors of about 1 mol / L of IC50 were identified by high throughput screening, and the homologous enzyme CSE was 180 times more selective. [conclusion] under the basic enzyme activity condition, the high throughput screening has found the CBS high efficiency natural product inhibitor whose selectivity is more than 180 times, which can provide the special molecular tool for the study of the molecular mechanism and function of CBS.
【作者单位】：上海交通大学系统生物医学教育部重点实验室系统生物医学研究院
【基金】：国家自然科学基金-青年科学基金项目(“靶向胱硫醚γ裂解酶抑制剂的发现及作用机制研究”,No.31500635)
【分类号】：R965.1

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