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Characterization of the histone methyltransferase PRDM9 using biochemical, biophysical and chemical biology techniques
journal contribution
posted on 2023-06-09, 19:35 authored by Xiaoying Koh-Stenta, Joma Joy, Anders Poulsen, Rong Li, Yvonne Tan, Yoonjung Shim, Jung-Hyun Min, Liling Wu, Anna Ngo, Jianhe Peng, Wei Guang Seetoh, Jing Cao, John Liang Kuan Wee, Perlyn Zekui Kwek, Alvin Hung, Umayal Lakshmanan, Horst Flotow, Ernesto Guccione, Jeffrey HillPRDM proteins have emerged as important regulators of disease and developmental processes. To gain insight into the mechanistic actions of the PRDM family, we have performed comprehensive characterization of a prototype member protein, the histone methyltransferase PRDM9, using biochemical, biophysical and chemical biology techniques. In the present paper we report the first known molecular characterization of a PRDM9-methylated recombinant histone octamer and the identification of new histone substrates for the enzyme. A single C321P mutant of the PR/SET domain was demonstrated to significantly weaken PRDM9 activity. Additionally, we have optimized a robust biochemical assay amenable to high-throughput screening to facilitate the generation of small-molecule chemical probes for this protein family. The present study has provided valuable insight into the enzymology of an intrinsically active PRDM protein.
History
Publication status
- Published
File Version
- Accepted version
Journal
Biochemical JournalISSN
0264-6021Publisher
Portland PressExternal DOI
Issue
2Volume
461Page range
323-334Department affiliated with
- Biochemistry Publications
Research groups affiliated with
- Sussex Drug Discovery Centre Publications
Full text available
- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2019-11-11First Open Access (FOA) Date
2019-11-11First Compliant Deposit (FCD) Date
2019-11-11Usage metrics
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