The semisynthesis of site-specifically modified histones and histone-based probes of chromatin-modifying enzymes

•Histone PTMs play an important role in regulating gene transcription.•Semisynthetic histones with defined PTMs are key to investigating the histone code.•Designer nucleosomes reveal specific biochemical and biophysical effects of PTMs.•Activity-based probes will allow identification of histone code...

Full description

Saved in:
Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 2023-07, Vol.215, p.28-37
Main Authors: Currie, Madeline F., Singh, Sumeet K., Ji, Meihuan, Chatterjee, Champak
Format: Article
Language:English
Subjects:
active sites
Activity-based probe
Chromatin - genetics
disulfides
enzyme substrates
enzymes
Histone
histones
Histones - metabolism
lysine
methylation
Nucleosome
nucleosomes
Nucleosomes - genetics
Post-translational modifications
Protein Processing, Post-Translational
Semisynthesis
sumoylation
transcription (genetics)
Ubiquitination
Ubiquitylation
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:•Histone PTMs play an important role in regulating gene transcription.•Semisynthetic histones with defined PTMs are key to investigating the histone code.•Designer nucleosomes reveal specific biochemical and biophysical effects of PTMs.•Activity-based probes will allow identification of histone code writers and erasers. Histone post-translational modifications (PTMs) on lysine residues, including methylation, ubiquitylation, and sumoylation, have been studied using semisynthetic histones reconstituted into nucleosomes. These studies have revealed the in vitro effects of histone PTMs on chromatin structure, gene transcription, and biochemical crosstalk. However, the dynamic and transient nature of most enzyme-chromatin interactions poses a challenge toward identifying specific enzyme-substrate interactions. To address this, we report methodology for the synthesis of two ubiquitylated activity-based probe histones, H2BK120ub(G76C) and H2BK120ub(G76Dha), that may be used to trap enzyme active-site cysteines as disulfides or in the form of thioether linkages, respectively. The general synthetic method we report for converting ubiquitylated nucleosomes into activity-based probes may also be applied to other histone sites of ubiquitylation in order to facilitate the identification of enzyme-chromatin interactions.
ISSN:1046-2023
1095-9130
1095-9130
DOI:10.1016/j.ymeth.2023.05.004