- API Development & Manufacture
- Peptide & Protein Technology
- rapidd™
- Analytical
- Solid State Services
- Isotope Chemistry
- Biocatalysis / selectAZyme™
- FLEXYTE™ Fluorescence Lifetime Assays
- Animal Free Chemokines
Keyword
Protein-Ligand Interactions
Epi-readers are proteins which recognise post-translational modifications on histones and play key roles in transcriptional regulation and chromatin remodelling. Examples include bromodomains which recognise acetylated lysine residues, and chromodomains and malignant-brain tumour domains which recognise methylated lysine residues. Such epi-readers are a major focus in drug discovery due to their role in oncology and inflammatory diseases
BRD4(1) Assay
- We have developed an assay for bromodomain, BRD4(1), whereby binding of a histone H4 acetylated peptide containing a FLT modulator residue, M, results in a change in fluorescence lifetime of 9AA labelled BRD4(1)
- Using our in-house protein engineering technologies, site-specifically 9AA labelled BRD4(1) was provided in good yield and excellent purity.
- Binding of 9AA labelled BRD4(1) bromodomain to an acetylated histone H4 peptide is reported through dose-dependent changes in FLT. The analogous unacetylated peptide does not cause a similar change in FLT confirming the ability of the system to report protein-ligand binding
- The FLEXYTE™ BRD4(1) assay was validated for inhibitor screening using the small molecule inhibitor, JQ1(+) and DMSO (an acetyl-lysine mimic) with IC50 values in good agreement with those reported in the literature
- A Z’-factor of 0.62 demonstrates the potential of the assay for HTS
- This platform technology can be readily developed for monitoring other protein-protein or protein-ligand interactions, as well as providing a platform technology for assaying Epi-reader proteins.
