收稿日期: 2016-03-21
网络出版日期: 2018-03-05
基金资助
国家科技部重大专项资助项目(2014ZX09507-002-004)
Establishment of molecular-based high-throughout screening model for histone lysine methyltransferase DOT1L inhibitors
Received date: 2016-03-21
Online published: 2018-03-05
类端粒沉默干扰体 1(disruptor of telomeric silencing 1-like, DOT1L) 是一种能够催化组蛋白 H3 第 79 位赖氨酸(H3K79)发生甲基 化修饰的表观遗传调控酶, 是第一个被发现不含有 SET 结构域的赖氨酸甲基转移酶. H3K79 位点的甲基化程度能够影响和调控某些特定基因的表达, 与急性髓性白血病(acute myelocytic leukemia, AML)的发生与发展密切相关. 为建立一种分子水平 DOT1L 小分子抑制剂的高通量筛选模型, 以 EPZ-5676 为阳性化合物, 探讨最佳反应酶浓度、底物浓度及反应时间等, 并对优化后的反应体系进行检测和确认. 通过对多种参数优化使得高通量筛选体系的 $Z'$ 因子达到 0.54, 表明已成功建立了 DOT1L 小分子抑制剂高通量筛选模型.
徐威, 苏明波, 周宇波 . 组蛋白赖氨酸甲基转移酶 DOT1L 抑制剂分子水平高通量筛选模型的建立[J]. 上海大学学报(自然科学版), 2018 , 24(1) : 126 -133 . DOI: 10.12066/j.issn.1007-2861.1757
DOT1L is an epigenetic enzyme that can catalyze methyltransfer activity at histone 3 lysine 79 (H3K79). It is also found to be the first lysine methyltransferase without a SET domain. Research results show that methylation level of H3K79 can influence the expression of some genes, closely related to the development of acute myelocytic leukemia (AML). This work establishes a molecular-based high-throughout screening model for discovering novel DOT1L inhibitors. EPZ-5676 was used as positive control to optimize activity assay and ensure appropriate concentration of enzyme and substrate, and reaction time, etc. The assay was optimized, and the $Z'$-factor was about 0.54, indicating that a molecular-based high-throughout screening model for DOT1L inhibitors was successfully established.
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