Histone Demethylase Screening: Identifying Inhibitors against HDMTs at Creative BioMart, Slides of Biology

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HDM Inhibitor Screening

The identification of histone demethylases (HDMT) catalyzing the removal of methyl groups from

modified histone proteins has indicated that demethylation events play an important role in

histone modification dynamics. Just like all other histone modifying enzymes, histone

demethylases play an important role in development and gene regulation, and many of them

require a cofactor like Fe(II), FAD, or α-ketoglutarate. HDMTs display a high level of substrate ketoglutarate. HDMTs display a high level of substrate

specificity to their target lysine or arginine, as well as high sensitivity to the specific type of

methylation to be recognized and demethylated. Currently, the most commonly studied histone

demethylases are LSD1 and JARIDs, where LSD1 can eliminate mono-ketoglutarate. HDMTs display a high level of substrate and di-ketoglutarate. HDMTs display a high level of substrate methylation from H3-ketoglutarate. HDMTs display a high level of substrate

K4 by using an amine oxidase reaction. The role of histone demethylases in the chromatin

regulation has provided new insight for understanding the epigenetic landscape of the chromatin

environment of cancer cells, thus making it necessary to better understand the activity or

inhibition of HDMTs and clarify their potential mechanisms in cancer therapy.

b. Fluorometrical Method

Fluorometrical method used by Creative BioMart allows a convenient measure of histone

demethylase inhibition, where the inhibitors directly interact with HDMT or block the binding of

the enzyme to its substrate. In the assay, HDMT is incubated with the unique substrate and

inhibitor in the strip wells, where the binding of enzyme with substrate produces hydrogen

peroxide. Then the fluorescent developing solution is added to react with the produced

hydrogen peroxide, thus obtaining the highly fluorescent oxidation products. The intensity of

fluorescence from oxidation products is proportional to enzyme activity. Therefore, as enzyme

activity decreases by inhibition, the fluorescent signal decreases.

Advantages of Our Service

Screening for multiple HDMTs targets or inhibitors

• IC

50

(half maximal inhibitory concentration) determination and single point concentration assay

Get project guidance in a time-efficient manner

Quick feedback data after sample submission

Fast procedure of fluorometrical method

Innovative fluorescent assay without need for radioactivity, extraction, and chromatography

Flexible strip microplate format for manual or high throughput analysis

Simple, reliable, and consistent assay conditions

Thank you!

Excerpt from:

https://www.creativebiomart.net/epigenetics/services/

epigenetic-drug-discovery/biochemical-assays/

demethylase-screening/histone-demethylase-screening/