Protocol for preparing SUV420H1 in complex with the nucleosome containing H2A.Z and H4K20Ecx for structure determination
The histone lysine methyltransferase SUV420H1 preferentially targets the H2A.Z-containing nucleosome core particle (H2A.Z-NCP) and catalyzes the H4K20me2 modification at replication origins. Here, we present a protocol for preparing SUV420H1 in complex with the nucleosome containing H2A.Z and H4K20Ecx for structure determination. We describe steps for the installation of S-ethyl-cysteine (Ecx), nucleosome and complex preparation, and performing the cryoelectron microscopy (cryo-EM) sample check. This protocol substitutes lysine 20 in histone H4 with S-ethyl-cysteine (H4K20Ecx), which enhances the stability of the interaction between SUV420H1 and nucleosomes. For complete details on the use and execution of this protocol, please refer to Huang et al.<sup>1</sup>.
Authors
- Zheng Zhou (ORCID: https://orcid.org/0000-0002-1423-7231)
- Li Huang (ORCID: https://orcid.org/0000-0003-2421-8162)
Institutions
- Institute of Biophysics (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-09-01
- DOI
- https://doi.org/10.1016/j.xpro.2024.103295
- Primary Topic
- RNA modifications and cancer
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- Ministry of Science and Technology of the People's Republic of China
- Institute of Biophysics, Chinese Academy of Sciences