Characterization of the histone methyltransferase PRDM9 using biochemical, biophysical and chemical biology techniques
Copyright policy )Characterization of the histone methyltransferase PRDM9 using biochemical, biophysical and chemical biology techniques
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.
- Experimental Drug Development Centre Singapore
- University of Illinois at Chicago United States
- Agency for Science, Technology and Research Singapore
- Biomedical Research Council Singapore
- Institute of Molecular and Cell Biology Singapore
Models, Molecular, Proline, Molecular Sequence Data, Gene Expression, Histone-Lysine N-Methyltransferase, Recombinant Proteins, High-Throughput Screening Assays, Protein Structure, Tertiary, Substrate Specificity, Histones, Kinetics, Mice, Luminescent Measurements, Mutation, Escherichia coli, Animals, Humans, Amino Acid Sequence, Cysteine, Protein Multimerization
Models, Molecular, Proline, Molecular Sequence Data, Gene Expression, Histone-Lysine N-Methyltransferase, Recombinant Proteins, High-Throughput Screening Assays, Protein Structure, Tertiary, Substrate Specificity, Histones, Kinetics, Mice, Luminescent Measurements, Mutation, Escherichia coli, Animals, Humans, Amino Acid Sequence, Cysteine, Protein Multimerization
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