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Issue 2, 2016
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Design of a fluorescent ligand targeting the S-adenosylmethionine binding site of the histone methyltransferase MLL1

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Abstract

The histone methyltransferase MLL1 has been linked to translocation-associated gene fusion in childhood leukemias and is an attractive drug target. High-throughput biochemical analysis of MLL1 methyltransferase activity requires the production of at least a trimeric complex of MLL1, RbBP5 and WDR5 to elicit robust activity. Production of trimeric and higher order MLL1 complexes in the quantities and reproducibility required for high-throughput screening presents a significant impediment to MLL1 drug discovery efforts. We present here a small molecule fluorescent ligand (FL-NAH, 6) that is able to bind to the S-adenosylmethionine (SAM) binding site of MLL1 in a manner independent of the associated complex members. We have used FL-NAH to develop a fluorescence polarization-based SAM displacement assay in a 384-well format targeting the MLL1 SET domain in the absence of associated complex members. FL-NAH competes with SAM and is displaced from the MLL1 SET domain by other SAM-binding site ligands with Kdisp values similar to the higher-order complexes, but is unaffected by the H3 peptide substrate. This assay enables screening for SAM-competitive MLL1 inhibitors without requiring the use of trimeric or higher order MLL1 complexes, significantly reducing screening time and cost.

Graphical abstract: Design of a fluorescent ligand targeting the S-adenosylmethionine binding site of the histone methyltransferase MLL1

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Publication details

The article was received on 26 Aug 2015, accepted on 29 Oct 2015 and first published on 29 Oct 2015


Article type: Paper
DOI: 10.1039/C5OB01794G
Citation: Org. Biomol. Chem., 2016,14, 631-638
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    Design of a fluorescent ligand targeting the S-adenosylmethionine binding site of the histone methyltransferase MLL1

    Y. Luan, L. L. Blazer, H. Hu, T. Hajian, J. Zhang, H. Wu, S. Houliston, C. H. Arrowsmith, M. Vedadi and Y. G. Zheng, Org. Biomol. Chem., 2016, 14, 631
    DOI: 10.1039/C5OB01794G

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