A fluorescence resonance energy transfer-based method for histone methyltransferases |
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| Authors: | Kanchan Devkota Brian Lohse Camilla Nyby Jakobsen Jens Berthelsen Rasmus Prætorius Clausen |
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| Institution: | 1. Department of Drug Design and Pharmacology, University of Copenhagen, DK-2100 Copenhagen, Denmark;2. NNF Center for Protein Research, University of Copenhagen, DK-2200 Copenhagen, Denmark;3. Department of International Health, Immunology and Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark |
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| Abstract: | A simple dye–quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye and a quencher, respectively. When lysine-9 residues in the peptides were methylated, they were protected from cleavage by endoproteinase–EndoLysC, whereas unmethylated peptides were cleaved, resulting in an increase in fluorescent intensity. |
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| Keywords: | Methyltransferase Assay EHMT1 |
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