Merck

Inhibition of histone binding by supramolecular hosts.

The Biochemical journal (2014-03-01)
Hillary F Allen, Kevin D Daze, Takashi Shimbo, Anne Lai, Catherine A Musselman, Jennifer K Sims, Paul A Wade, Fraser Hof, Tatiana G Kutateladze
ABSTRACT

The tandem PHD (plant homeodomain) fingers of the CHD4 (chromodomain helicase DNA-binding protein 4) ATPase are epigenetic readers that bind either unmodified histone H3 tails or H3K9me3 (histone H3 trimethylated at Lys⁹). This dual function is necessary for the transcriptional and chromatin remodelling activities of the NuRD (nucleosome remodelling and deacetylase) complex. In the present paper, we show that calixarene-based supramolecular hosts disrupt binding of the CHD4 PHD2 finger to H3K9me3, but do not affect the interaction of this protein with the H3K9me0 (unmodified histone H3) tail. A similar inhibitory effect, observed for the association of chromodomain of HP1γ (heterochromatin protein 1γ) with H3K9me3, points to a general mechanism of methyl-lysine caging by calixarenes and suggests a high potential for these compounds in biochemical applications. Immunofluorescence analysis reveals that the supramolecular agents induce changes in chromatin organization that are consistent with their binding to and disruption of H3K9me3 sites in living cells. The results of the present study suggest that the aromatic macrocyclic hosts can be used as a powerful new tool for characterizing methylation-driven epigenetic mechanisms.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Lysine monohydrochloride, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Lysine monohydrochloride, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Lysine monohydrochloride, BioUltra, ≥99.5% (AT)
Supelco
L-Lysine monohydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
L-Lysine Acetate, Pharmaceutical Secondary Standard; Certified Reference Material
Lysine hydrochloride, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
L-Lysine monohydrochloride, Vetec, reagent grade, ≥98%
Sigma-Aldrich
Nε,Nε,Nε-Trimethyllysine hydrochloride, ≥97% (TLC)
Supelco
L-Lysine monohydrochloride, certified reference material, TraceCERT®
Sigma-Aldrich
Anti-trimethyl-Histone H3 (Lys9) Antibody, serum, Upstate®
Sigma-Aldrich
L-Lysine, crystallized, ≥98.0% (NT)
Sigma-Aldrich
L-Lysine, Vetec, reagent grade, ≥98%
Lysine acetate, European Pharmacopoeia (EP) Reference Standard
Supelco
L-Lysine, analytical standard
Sigma-Aldrich
L-Lysine, ≥98% (TLC)
Sigma-Aldrich
L-Lysine acetate salt, ≥98% (HPLC)