以活动为导向的化甲基素摄影捕获用于化甲基酶的基质剖析
Jordan Kuwik1, Kathryn Hinkelman1, Megan Waldman1
1Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.
Journal of the American Chemical Society
|September 13, 2023
概括
研究人员开发了一种新的方法,即交联辅助基质识别 (CASI),以识别移转酶的细胞点,这些酶可以去除修饰. 这种方法成功地绘制了氨酸脱甲基酶 (KDM) 的新点,揭示了基因转录以外的更广泛的作用.
科学领域:
- 生物化学
- 蛋白质组学
- 分子生物学
背景情况:
- 可逆转化后修饰 (PTM) 通过蛋白相互作用调节细胞信号.
- 转移酶去除PTM,但使用当前的方法很难识别它们的基质.
- 由于表皮质损失,现有的化学蛋白质技术对转移酶基质的分类无效.
研究的目的:
- 为系统地识别转移酶的目标制定一个一般的化学蛋白质策略.
- 将这种策略应用于 lysine 脱甲基酶 (KDM) 并发现它们的细胞点.
- 扩大对KDM功能的理解,而不仅仅是它们的已知作用.
主要方法:
- 开发了交叉链接辅助基质识别 (CASI),是一种新的化学蛋白质战略.
- 用p-azido氨酸对KDM4进行向光反应.
- 使用CASI进行KDM基板的蛋白质组测量.
主要成果:
- 发现了一种新的KDM细胞点"脱甲基".
- 发现了KDM4的众多非基质,扩大了其已知的生物功能.
- 发现KDM4A在转化启动因子中的关键残留物去甲基化,这表明它在核糖体过程中的作用.
结论:
- CASI是一种强大的新方法来识别转移酶基质,克服了以前方法的局限性.
- KDM4A在细胞过程中具有比以前更广泛的作用,包括翻译.
- 可以将CASI应用于400多个人类转移酶,从而促进对鲜为人知的酶的研究.
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