Koh-Stenta, Xiaoying, Joy, Joma, Poulsen, Anders, Li, Rong, Tan, Yvonne, Shim, Yoonjung, Min, Jung-Hyun, Wu, Liling, Ngo, Anna, Peng, Jianhe, Seetoh, Wei Guang, Cao, Jing, Wee, John Liang Kuan, Kwek, Perlyn Zekui, Hung, Alvin, Lakshmanan, Umayal, Flotow, Horst, Guccione, Ernesto and Hill, Jeffrey (2014) Characterization of the histone methyltransferase PRDM9 using biochemical, biophysical and chemical biology techniques. Biochemical Journal, 461 (2). pp. 323-334. ISSN 0264-6021

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Abstract

PRDM 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.

Item Type:	Article
Schools and Departments:	School of Life Sciences > Biochemistry
Research Centres and Groups:	Sussex Drug Discovery Centre
Depositing User:	Jeffrey Hill
Date Deposited:	11 Nov 2019 08:32
Last Modified:	11 Nov 2019 11:45
URI:	http://sro.sussex.ac.uk/id/eprint/87935

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