Histone methyltransferases (HMTs) that catalyze histone methylation could control or regulate DNA methylation through chromatin-dependent transcription repression or activation. Histone methylation participates in both epigenetic gene activation and silencing, thus making it important to measure the activity or inhibition of HMTs. In particular, methylation of histone 3 at lysine 4, lysine 9, and lysine 27 are implicated in some pathological processes, making HMT excellent targets for pharmacological intervention. There is a growing interest in identifying corresponding small molecule inhibitors with therapeutic potential.

Creative BioMart launches small molecule screening campaigns for HMTs in their native cellular state, thus providing a much more biologically relevant assessment of inhibitor effects on epigenetic marks as well as revealing the cell permeability and toxicity of inhibitors. These screening services are supported by AlphaLisa assay and Western blot assay platforms. The implementation of these cell-based assays will positively impact your discovery program of pharmacologically potent HMT inhibitors. Creative BioMart also offers our customers commercial cell lines, ready-to-use assay kits and tool-box reagents.

Procedure of Cell-based AlphaLisa Assays at Creative BioMart

Cells are seeded to assay plates and incubated with compound for a few days at appropriate temperature in CO₂. A sample containing biotinylated peptide substrate and HMT is then added to the testing system. Subsequently, acceptor beads and donor beads are added, followed by reading the Alpha response. Finally, the IC₅₀ (half maximal inhibitory concentration) curves are plotted based on a sigmoidal dose-response equation and IC₅₀ values of tested compounds are calculated to evaluation of the cellular inhibitory activity. This AlphaLisa sandwich assay has advantages of high sensitivity, dynamic range, scalability and resistance to fluorescent interference.

Figure 1. Schematic procedure of cell-based histone methylation assay

Several different cell based histone methylation assays are accessible at Creative BioMart for your early biopharmaceutical research. Owing to the diversity of histone methylation sites and patterns, the selection of an appropriate assay platform for screening and profiling against HMT requires special considerations. Our team of experienced molecular, assay and cell biologists is able to guide you to choose the most suitable tools and assist you in a way precisely adapted to your needs. Whether you have specific requirements or are looking for a partner to provide a full range of support, please feel free to contact us!

Reference
1. Martinez, N. J.; Simeonov, A. Cell-based assays to support the profiling of small molecules with histone methyltransferase and demethylase modulatory activity. Drug Discov Today Technol. 2015, 18: 9-17.