Abstract
We describe the discovery of UNC1215, a potent and selective chemical probe for the methyllysine (Kme) reading function of L3MBTL3, a member of the malignant brain tumor (MBT) family of chromatin-interacting transcriptional repressors. UNC1215 binds L3MBTL3 with a K(d) of 120 nM, competitively displacing mono- or dimethyllysine-containing peptides, and is greater than 50-fold more potent toward L3MBTL3 than other members of the MBT family while also demonstrating selectivity against more than 200 other reader domains examined. X-ray crystallography identified a unique 2:2 polyvalent mode of interaction between UNC1215 and L3MBTL3. In cells, UNC1215 is nontoxic and directly binds L3MBTL3 via the Kme-binding pocket of the MBT domains. UNC1215 increases the cellular mobility of GFP-L3MBTL3 fusion proteins, and point mutants that disrupt the Kme-binding function of GFP-L3MBTL3 phenocopy the effects of UNC1215 on localization. Finally, UNC1215 was used to reveal a new Kme-dependent interaction of L3MBTL3 with BCLAF1, a protein implicated in DNA damage repair and apoptosis.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Benzamides / chemistry
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Benzamides / metabolism
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Benzamides / pharmacology*
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Binding, Competitive / drug effects
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Crystallography, X-Ray
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DNA-Binding Proteins / antagonists & inhibitors
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / metabolism*
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Dose-Response Relationship, Drug
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Drug Discovery*
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HEK293 Cells
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Humans
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Lysine / analogs & derivatives*
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Lysine / antagonists & inhibitors
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Lysine / chemistry
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Lysine / metabolism
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Models, Molecular
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Molecular Probes / chemistry
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Molecular Probes / metabolism
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Molecular Probes / pharmacology*
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Molecular Structure
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Piperidines / chemistry
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Piperidines / metabolism
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Piperidines / pharmacology*
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Protein Structure, Tertiary
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Repressor Proteins / metabolism
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Structure-Activity Relationship
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Tumor Suppressor Proteins / metabolism
Substances
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BCLAF1 protein, human
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Benzamides
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DNA-Binding Proteins
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L3MBTL3 protein, human
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Molecular Probes
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Piperidines
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Repressor Proteins
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Tumor Suppressor Proteins
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UNC1215
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2-methyllysine
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Lysine
Associated data
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PDB/4FL6
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PubChem-Substance/160647817
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PubChem-Substance/160647818
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PubChem-Substance/160647819
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PubChem-Substance/160647820
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PubChem-Substance/160647821