lsd1 screening assay kit  (Cayman Chemical)

Journal: Molecules

Article Title: Identification and Evaluation of Olive Phenolics in the Context of Amine Oxidase Enzyme Inhibition and Depression: In Silico Modelling and In Vitro Validation

doi: 10.3390/molecules29112446

Figure Lengend Snippet: Molecular docking results for LSD1. ( A ) The crystal structure of LSD1 in complex with CoREST (PDB ID: 5YJB) is depicted. The FAD co-factor is labelled and predicted ligand-binding sites are coloured brown. 4-[5-(piperidin-4-ylmethoxy)-2-( p -tolyl)pyridin-3-yl]benzonitrile was used as the positive control inhibitor and was docked to the substrate-binding cavity of LSD1. The binding affinity was predicted to be −9.5 kcal/mol. ( B ) The phenolic compounds OLP, OLC, HTA, and HT were screened against the substrate-binding cavity and the binding affinities are provided (kcal/mol). Key residues are labelled, with those predicted to form hydrogen bonds and π–π stacking interactions italicised.

Article Snippet: The inhibitory activity of TCP, HT, HTA, and OLP against purified LSD1 was measured using a commercially available histone demethylase LSD1 inhibitory screening assay core kit (Epigentek P-3075A, Farmingdale, NY, USA) according to the manufacturer’s instructions.

Techniques: Ligand Binding Assay, Positive Control, Binding Assay

Journal: Molecules

Article Title: Identification and Evaluation of Olive Phenolics in the Context of Amine Oxidase Enzyme Inhibition and Depression: In Silico Modelling and In Vitro Validation

doi: 10.3390/molecules29112446

Figure Lengend Snippet: Protein–peptide docking results for histone H3 and LSD1. ( A ) Crystal structure of LSD1-CoREST in complex with the N-terminal residues of the histone H3 peptide. The FAD co-factor is coloured brown. Blind protein–peptide docking was performed to examine the preferential binding site of the histone H3 peptide in the presence of ligands bound to the substrate-binding cavity. The histone H3 peptide was docked to the crystal structure of LSD1-CoREST in the ( B ) absence and ( C ) presence of phenolic compounds bound to the substrate-binding cavity. The results are shown for OLC. ( Aii – Cii ) The protein–peptide interactions were evaluated using PDBePISA. The residues of the LSD1 substrate-binding cavity that were predicted to form salt bridges with R2 and R8 (underlined) of the histone H3 peptide are labelled.

Article Snippet: The inhibitory activity of TCP, HT, HTA, and OLP against purified LSD1 was measured using a commercially available histone demethylase LSD1 inhibitory screening assay core kit (Epigentek P-3075A, Farmingdale, NY, USA) according to the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Molecules

Article Title: Identification and Evaluation of Olive Phenolics in the Context of Amine Oxidase Enzyme Inhibition and Depression: In Silico Modelling and In Vitro Validation

doi: 10.3390/molecules29112446

Figure Lengend Snippet: Inhibitory activity of phenolic compounds against LSD1. ( A ) The results of the control compound TCP and the phenolic compounds HT, HTA, and OLP from the direct enzymatic assays are provided. The data presented denote the mean ± SEM from duplicate (TCP, HTA, and OLP) and triplicate (HT) assays (representative results from n = 3 independent experiments). ( B ) A separate set of experiments were performed by Reaction Biology Corporation using the positive control inhibitor HCl 489479 and the phenolic compounds OLP, HT, HTA, and OLC. The demethylase activity (%) of LSD1 was measured for the control compound HCl 489479 and the phenolic compounds at concentrations ranging from 0–10 μM and 0–100 μM, respectively.

Article Snippet: The inhibitory activity of TCP, HT, HTA, and OLP against purified LSD1 was measured using a commercially available histone demethylase LSD1 inhibitory screening assay core kit (Epigentek P-3075A, Farmingdale, NY, USA) according to the manufacturer’s instructions.

Techniques: Activity Assay, Control, Positive Control