Title Identification of a small-molecule ligand of the epigenetic reader protein Spindlin1 via a versatile screening platform /
Authors Wagner, Tobias ; Greschik, Holger ; Burgahn, Teresa ; Schmidtkunz, Karin ; Schott, Anne-Kathrin ; McMillan, Joel ; Baranauskienė, Lina ; Xiong, Yan ; Fedorov, Oleg ; Jin, Jian ; Oppermann, Udo ; Matulis, Daumantas ; Schule, Roland ; Jung, Manfred
DOI 10.1093/nar/gkw089
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Is Part of Nucleic acids research.. Oxford : Oxford University Press. 2016, Vol. 44, 9, p. 1-22.. ISSN 0305-1048. eISSN 1362-4962
Keywords [eng] Epigenetic modifications ; histone ; eukaryotic transcription ; epigenetic signalling ; ligands ; smallmolecule inhibitors.
Abstract [eng] Epigenetic modifications of histone tails play an essential role in the regulation of eukaryotic transcription. Writer and eraser enzymes establish and maintain the epigenetic code by creating or removing posttranslational marks. Specific binding proteins, called readers, recognize the modifications and mediate epigenetic signalling. Here, we present a versatile assay platform for the investigation of the interaction between methyl lysine readers and their ligands. This can be utilized for the screening of smallmolecule inhibitors of such protein–protein interactions and the detailed characterization of the inhibition. Our platform is constructed in a modular way consisting of orthogonal in vitro binding assays for ligand screening and verification of initial hits and biophysical, label-free techniques for further kinetic characterization of confirmed ligands. A stability assay for the investigation of target engagement in a cellular context complements the platform. We applied the complete evaluation chain to the Tudor domain containing protein Spindlin1 and established the in vitro test systems for the double Tudor domain of the histone demethylase JMJD2C. We finally conducted an exploratory screen for inhibitors of the interaction between Spindlin1 and H3K4me3 and identified A366 as the first nanomolar small-molecule ligand of a Tudor domain containing methyl lysine reader.
Published Oxford : Oxford University Press
Type Journal article
Language English
Publication date 2016