Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong, China.
STAR Protoc. 2021 Jun 12;2(3):100604. doi: 10.1016/j.xpro.2021.100604. eCollection 2021 Sep 17.
Lysine succinylation is a recently discovered posttranslational modification that plays critical roles in metabolism, epigenetic signaling, and human diseases. To investigate the effects of site-specific histone lysine succinylation on nucleosome dynamics requires the generation of homogeneously modified histones, which is a significant challenge. Here, we report a protocol for the rapid site-specific installation of a succinyl lysine analog onto histone. We then use a Förster resonance energy transfer approach to characterize the impact on nucleosome dynamics. For complete details on the use and execution of this protocol, please refer to Jing et al. (2018).
赖氨酸琥珀酰化是一种新发现的翻译后修饰,在代谢、表观遗传信号和人类疾病中发挥着关键作用。为了研究组蛋白赖氨酸特异性琥珀酰化对核小体动力学的影响,需要生成均一地修饰的组蛋白,这是一个重大挑战。在这里,我们报告了一种在组蛋白上快速特异性安装琥珀酰化赖氨酸类似物的方案。然后,我们使用Förster 共振能量转移方法来表征其对核小体动力学的影响。有关该方案的使用和执行的完整详细信息,请参阅 Jing 等人(2018 年)。