对于对模拟敏感的TET氧基酶的蛋白质 - 连接器接口的补充固体工程
Babu Sudhamalla1, Sinan Wang1, Valerie Snyder1
1Department of Chemistry , University of Pittsburgh , Pittsburgh , Pennsylvania 15260 , United States.
Journal of the American Chemical Society
|July 21, 2018
概括
研究人员设计了十一转位 (TET) 酶以创建对模拟敏感的变体. 这允许特异性抑制TET2,使得人类细胞中DNA细胞酶甲基化的条件调节.
科学领域:
- 生物化学
- 分子生物学
- 表观遗传学
背景情况:
- 十一转位 (TET) 酶对于通过DNA脱甲基的哺乳动物基因组调节至关重要.
- 人体中存在60多种C-H氧酶,因此很难确定单个酶的功能.
- 针对特定的TET酶对于理解它们在细胞过程中的作用至关重要.
研究的目的:
- 开发一种专门扰乱单个TET酶活动的方法.
- 设计用于条件抑制的模拟敏感的TET2变体.
- 建立一个研究2基酸盐 (2KG) 依赖氧化酶的一般平台.
主要方法:
- 合理设计TET2-2-酸盐 (2KG) 接口以创建具有扩展活性位点的变体.
- 对TET蛋白质的一种庞大的守门者残留物的鉴定和工程.
- 使用N-oxalylglycine (NOG) 衍生物开发直角突变抑制剂对.
- 在完整的人类细胞中应用细胞透的NOG类似物来抑制TET2突变.
主要成果:
- 成功设计了对特定NOG衍生物敏感的TET2变种.
- 证明了TET2突变的正交抑制,允许条件调节细胞因子甲基化.
- 展示了其他TET蛋白的方法的普遍性.
- 应用该系统来探测细胞中TET介导的转录活性.
结论:
- 开发了一个多功能平台,用于创建模拟敏感的2KG依赖氧酶.
- 能够精确控制TET酶的活性,以研究它们在各种信号通路中的功能.
- 在基因组调节和细胞过程中剖析特定的C-H氧酶的作用提供了一个新工具.
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