Thermodynamic analyses of Sgf29-H3K4me2 and Sgf29-H3K4me3 interactions.

<p>Representative ITC experiments showing the titration of H3K4me3 (top row, A-D) and H3K4me2 (bottom row, E-H) peptides to Sgf29 (first column, A,E), D266E (second column B, F), D266N (third column, C, G) and D266Y (fourth column, D, F).</p

Tm values for Sgf29 and its variants as determined by DSF.^[a]

<p>^[a] Tm as measured by DSF ± SD (Measured in triplicate)</p><p>Tm values for Sgf29 and its variants as determined by DSF.<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0139205#t001fn001" target="_blank">^[a]</a></p

Tm curves of A) wild-type Sgf29 and its D266 variants and B) Sgf29 and its Y238 and Y245 variants.^[a]

<p>Tm curves of A) wild-type Sgf29 and its D266 variants and B) Sgf29 and its Y238 and Y245 variants.^[a]</p

CD spectra of various expressions of wild-type Sgf29 its D266 variants, and the Y238F and Y245F variants.

<p>CD experiments were carried out at the concentration of 0.1 mg ml^-1 in 10 mM sodium phosphate buffer (pH 7.5).</p

Representative ITC experiments displaying binding of H3K4me3 and H3K4me2 to Y238F and Y245F Sgf29 variants.

<p>A) H3K4me3-Y238F, B) H3K4me3-Y245F, C) H3K4me2-Y238F and D) H3K4me2-Y245F.</p

ITC experiments showing binding of A) ARTKme3QTAGKS and B) ARTKme3QTA to WT Sgf29.

<p>Thermodynamics of binding for A) <i>K</i>_d = 4.0 ± 0.6 μM, Δ<i>G</i>° = - 7.4 ± 0.1 kcal mol^-1, Δ<i>H</i>° = - 8.0 ± 0.1 kcal mol^-1,-TΔ<i>S</i>° = 0.6 ± 0.1 kcal mol^-1 and for B) <i>K</i>_d = 9.0 ± 0.5 μM, Δ<i>G</i>° = - 6.9 ± 0.1 kcal mol^-1, Δ<i>H</i>° = - 7.8 ± 0.1 kcal mol^-1,-TΔ<i>S</i>° = 0.9 ± 0.1 kcal mol^-1.</p

Crystal structure of Sgf29-H3K4me3 (A, PDB ID: 3ME9), Sgf29-H3K4me2 (B, PDB ID: 3MET) and the apo form of Sgf29 (C, PDB ID: 3MEW).

<p>Sgf29 and H3K4me2/3 histone peptide are shown in cyan and yellow, respectively.</p

Investigations on Small Molecule Inhibitors Targeting the Histone H3K4 tri-methyllysine Binding PHD-finger of JmjC Histone Demethylases

Plant homeodomain (PHD) containing proteins are important epigenetic regulators and are of interest as potential drug targets. Inspired by the amiodarone derivatives reported to inhibit the PHD finger 3 of KDM5A (KDM5A(PHD3)), a set of compounds were synthesised. Amiodarone and its derivatives were observed to weakly disrupt the interactions of a histone H3K4me3 peptide with KDM5A(PHD3). Selected amiodarone derivatives inhibited catalysis of KDM5A, but in a PHD-finger independent manner. Amiodarone derivatives also bind to H3K4me3-binding PHD-fingers from the KDM7 subfamily. Further work is required to develop potent and selective PHD finger inhibitors

Human histone demethylase KDM6B can catalyse sequential oxidations

Jumonji domain‐containing demethylases (JmjC‐KDMs) catalyse demethylation of Nε ‐methylated lysines on histones and play important rolesin gene regulation. We report selectivity studies on KDM6B (JMJD3), a disease‐relevant JmjC‐KDM, using synthetic lysine analogues. The results unexpectedly reveal that KDM6B accepts multiple Nε ‐alkylated lysine analogues, forming alcohol, aldehyde and carboxylic acid products