全球甲基化抑制剂3-Deazaneplanocin A (DZNep) 诱导的蛋白质热稳定性的变化
Christine A Berryhill1, Emma H Doud1,2,3, Jocelyne N Hanquier1
1Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Biomolecules
|July 27, 2024
概括
DZNep治疗改变了蛋白质的热稳定性,影响了135种蛋白质,包括许多氨酸甲基化蛋白质. 这项研究揭示了DZNep.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- DZNep (3-deazaneplanocin A) 是已知的S-adenosyl-l-homocysteine水解酶 (AHCY) 抑制剂,通过改变SAM-to-SAH比率来影响细胞甲基化潜力.
- 之前的研究重点是DZNep对基因组 lysine甲基化的影响,对其对非基因组蛋白更广泛的影响的理解有限.
- 蛋白质热稳定性测试提供了一种新的方法来研究小分子抑制剂的作用机制,反映了连接物结合,PTM和蛋白质相互作用.
研究的目的:
- 描述DZNep处理对HEK293T细胞中蛋白质热稳定性的下游影响.
- 调查转录和蛋白质丰度的变化,以区分对热稳定的直接和间接影响.
- 探索DZNep诱导的热稳定性变化与蛋白质的氨酸甲基化状态之间的关系.
主要方法:
- 使用蛋白质整体可溶性变化 (PISA) 试验来测量DZNep治疗后蛋白质热稳定性的变化.
- 进行了转录组和蛋白质组范围的分析,以评估基因和蛋白质表达水平的变化.
- 分析特定蛋白质,如CDK6和甲基转移酶 (例如DNMT1),以检测热稳定性和甲基化状态的变化.
主要成果:
- DZNep治疗显著改变了135种蛋白质的热稳定性,其中超过一半被确定为氨酸甲基化.
- 大约三分之一的蛋白质表现出变化的热稳定性,在转录或蛋白质丰度水平上没有相应的变化.
- 在CDK6中观察到甲基化的稳定,而其非甲基化对应物保持不变;多个甲基转移酶,包括DNMT1,也显示了变化的热稳定性.
结论:
- DZNep 诱导了蛋白质热稳定性的广泛变化,超出了它已知的对基质子的影响,这表明它具有更广泛的细胞影响.
- 蛋白质热稳定性的改变可以独立于转录或蛋白质丰度的变化发生,突出显示后翻译性修改是关键驱动因素.
- 这项研究提供了对DZNep对转录组,蛋白质组和蛋白质热稳定性的系统评估,为其作用机制提供了新的见解.
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