Protocol for the preparation of site-specific succinylated histone mimics to investigate the impact on nucleosome dynamics

Abstract

Summary: 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).

Keywords

• Biophysics
• Molecular Biology
• Molecular/Chemical Probes
• Protein Biochemistry
• Protein expression and purification
• Chemistry