Proteins containing the methyl-DNA binding SET and RING-associated (SRA) domain are required for the establishment and/or maintenance of DNA methylation in both mammals and plants. The SRA domains of mammals and plants use both common and different recognition principles to target their DNA duplex sites. The SRA-containing mammalian UHRF1 protein plays a key role in epigenetic inheritance of 5-methylcytosines (5mC) marks in a 5mCpG context.

Aberrant DNA methylation may be prevented by small molecules preventing the interaction between the SRA domain and hemimethylated DNA. Therefore, the SRA domain of UHRF1 is emerging as an attractive target for the treatment of cancer characterized by UHRF1 overexpression and DNA hypermethylation. Small molecules binders of SRA domain of UHRF1 would be useful components of molecular toolbox for studying cancer epigenetics, cellular signaling pathways, and most notably as lead molecules for the development of anti-cancer therapeutics.

Figure 1. Mechanism of Small Molecule Binders of SRA Domain of UHRF1. (Patnaik D.; et al. 2018).

Creative BioMart provides the high-throughput screening assay based on time-resolved fluorescence resonance energy transfer (TR-FRET) to identify small molecule inhibitors that disrupt the interaction between the SRA domain and hemimethylated DNA.

SRA Domain Screening Assay at Creative BioMart

TR-FRET, which includes both HTRF and LANCE technology, is well documented as a sensitive and reliable method for the HTS of diverse enzyme and receptor targets. Typical TR-FRET immunoassays involve the homogeneous detection of biotinylated substrate or biotinylated product by the binding of europium (Eu) donor-tagged antibodies. These antibodies bring the donor molecule in close enough proximity so that when it is excited at 337 nm, its subsequent emission (590 nm for the Eu-cryptate, which is used in HTRF, or 620 nm for the Eu-chelate used in LANCE) can be absorbed by an appropriate streptavidin-tagged acceptor molecule (allophycocyanin or ULight), which then emits light at 665 nm.

Advantages of Choosing Creative BioMart

1. First-class technology platform: with the high-throughput compound screening technology platform, Creative BioMart is committed to cooperate with the extensive field to accelerate the discovery of epigenetic drugs.
2. One-stop & Customized service: you only need to provide us with samples or illustrate your research objectives, and we can screen inhibitors of SRA domain binding to hemimethylated DNA according to your specific research needs.
3. High-quality data: we perform quality control at every step of the procedure to ensure data repeatability and reliability, and we can validate hit compounds in a dose-response series.

Figure 2. Workflow of Our Service at Creative BioMart

Please don’t hesitate to contact us for more information about our SRA domain screening service or other services about epigenetic drug discovery.

References
1. Patnaik D.; et al. Targeting the SET and RING-associated (SRA) domain of ubiquitin-like, PHD and ring finger-containing 1 (UHRF1) for anti-cancer drug development. Oncotarget. 2018, 9(40): 26243-26258.
2. Yu V.; et al. High-throughput TR-FRET assays for identifying inhibitors of LSD1 and JMJD2C histone lysine demethylases. Journal of Biomolecular Screening. 2012, 17(1): 27.