Demethylase Assay Services

Creative Biolabs is the leading company offering demethylase assay services. With years of experience, Creative Biolabs provides access to a range of demethylase assay platforms along with affordable high throughput screening services.

Interest in the biological significance of lysine and arginine side-chain methylation and demethylation is growing in the fields of basic research and pharmacology. The dynamic methylation of histone H3 and H4 protein tails, in conjunction with other post-translational modifications, is a crucial process in the transcription control machinery. Numerous genetic connections have been found between illnesses and the catalytic domains responsible for altering the histone tail in both developing and adult organisms. Research into the corresponding demethylases (KDMs) has been sparked by the histone deacetylase inhibitors' clinical effectiveness. There are two families of KDMs that have been identified: the larger family of 2-oxoglutarate (2OG), ferrous iron, and oxygen-dependent demethylases (JmjC KDMs) and the flavin-dependent lysine-specific demethylases (LSDs or KDM1s).

Fig.1. Demethylation by Histone Demethylases.¹

Targets of Demethylase Assay Services

There are two families of KDMs that have been identified: KDM1s and JmjC KDMs. Both families catalyze N-methyl-lysine demethylation by oxidative processes. As members of the amine oxidase superfamily, the LSD enzymes (KDM1A/B) connect flavin adenine dinucleotide reduction to substrate oxidation. As members of the 2OG- and ferrous iron-dependent oxygenase (2OG oxygenase) superfamily, the JmjC KDMs catalyze the oxidative decarboxylation of 2OG to produce a highly reactive iron(IV)-oxo species. This species acts to hydroxylate the N-methyl group, forming an unstable hemi-aminal that breaks down to produce formaldehyde and the demethylated product. Creative Biolabs' demethylase assay services aim to achieve the following goals:

Case Studies of Small-Molecule Demethylase Inhibitor Discovery

• KDM1A

H3K4me2/1 is demethylated by KDM1A. KDM1A has been linked to both neurodevelopment and cancer. The MAO-inhibitor TCP is a mechanism-based, time-dependent KDM1A inactivator. Although the chemical is fundamentally an MAO inhibitor and its clinical usage as a KDM1A inhibitor has been proposed, it is preferable to remove its MAO-A inhibitory ability since it can lead to undesired and potentially fatal drug-drug and food-drug interactions.

• KDM4

Major H3K9me2/3 or H3K36me3 demethylation is done by the KDM4 subfamily. It consists of five members who have been connected to multiple cancer types. Key players in androgen signaling and putative prostate cancer progression factors are KDM4A/KDM4B. In squamous cell carcinoma, KDM4A is a determinant of invasiveness and metastasis, while KDM4B is substantially expressed in breast cancer that is ER+. In many hematological and solid malignancies, KDM4C is overexpressed and amplified. It was discovered that SD70, an 8-hydroxyquinoline derivative, inhibits KDM4C. Both in vivo (10 mg/kg/day) and in vitro (IC₅₀ 5–10 mM), the substance suppresses the development of prostate cells.

Creative Biolabs provides demethylase assay services that test compounds against demethylases for drug discovery. We provide a wide range of demethylases on our comprehensive platform to meet all of your research requirements. Expert researchers with an abundance of Creative Biolabs expertise will identify the best activators or inhibitors for your target demethylases. Please contact us for more information.

Reference

1. Maes, Tamara, et al. "Advances in the development of histone lysine demethylase inhibitors." Current opinion in pharmacology 23 (2015): 52-60.