Docket #: S19-163
High-Throughput Screening for Chemical Inhibitors of Histone Acetyltransferase
Histone acetyltransferase 1 (HAT1) is an enzyme which acetylates lysine on histone proteins and is intricately involved with regulating gene transcription. Thus, HAT1 is a promising therapeutic target for cancer and a high-throughput method to discover potent inhibitors would be valuable. A fluorescence-based, enzymatic assay can be constructed using a biotinylated HAT1 substrate, recombinant HAT1, and an acetyl co-A substrate mimetic with an alkyne click handle. The HAT1 substrate can be captured by wells coated with streptavidin. Biotin-azide can be conjugated to alkyne-containing lysine through click chemistry. Addition of streptavidin-HRP facilitates the measurement of activity via plate reader.
Credit: Permission from authors
Applications
- Fluorescence-based screening of novel HAT1 inhibitors on 96-well plates
- High-throughput screen design can be readily adapted to other acetyltransferases
Advantages
- No chemical HAT1 inhibitors currently exist, and this approach enables the discovery of potent HAT1 inhibitors
- High-throughput approach enables rapid measurement of enzymatic activity in vitro
- Plate reader approach makes it an affordable method
- No biological material necessary
- High dynamic range and sensitivity
Publications
- Gruber JJ, Geller B, Lipchik AM, Chen J, Salahudeen AA, Ram AN, Ford JM, Kuo CJ, Snyder MP. HAT1 Coordinates Histone Production and Acetylation via H4 Promoter Binding Mol Cell 2019 Aug 22;75(4):711-724.e5. . Epub 2019 Jul 2.
Related Links
Patents
- Published Application: 20220196660
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