Photoelectrochemical determination of the activity of histone acetyltransferase and inhibitor screening by using MoS2 nanosheets
The enzyme histone acetyltransferase (HAT) catalyzes the acetylation of a substrate peptide, and acetyl coenzyme A is converted to coenzyme A (CoA). A photoelectrochemical method is described for the determination of the HAT activity by using exfoliated MoS2 nanosheets, phos-tag-biotin, and β-galactosidase (β-Gal) based signal amplification. The MoS2 nanosheets are employed as the photoactive material, graphene nanosheets as electron transfer promoter, gold nanoparticles as recognition and capture reagent for CoA, and phos-tag-biotin as the reagent to link CoA and β-Gal. The enzyme β-Gal catalyzes the hydrolysis of substrate O-galactosyl-4-aminophenol to generate free 4-aminophenol which is a photoelectrochemical electron donor. The photocurrent increases with the activity of HAT. Under optimal conditions, the response is linear in the 0.3 to 100 nM activity range, and the detection limit is 0.14 nM (at S/N = 3). The assay was applied to HAT inhibitor screening, specifically for the inhibitors C646 and anacardic acid. The IC50 values are 0.28 and 39 μM, respectively. The method is deemed to be a promising tool for epigenetic research and HAT-targeted cancer drug discovery.
KeywordsPhos-tag-biotin Photoelectrochemistry C646 Anacardic acid 4-Aminophenol β-Galactosidase Graphene Acetyl coenzyme A Gold nanoparticles Visible light excitation
This work was supported by the National key research and development project of China (2018YFC1800605), the National Natural Science Foundation of China (No. 21775090), the Natural Science Foundation of Shandong province, China (No. ZR2016BM10), the Special Funds of Taishan Scholar of Shandong Province, China.
Compliance with ethical standards
The authors declare that they have no competing interests.
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