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Photoelectrochemical determination of the activity of histone acetyltransferase and inhibitor screening by using MoS2 nanosheets

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Abstract

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.

Histone acetyltransferase was detected using a sensitive photoelectrochemical method using MoS2 nanosheets as photoactive material.

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Acknowledgements

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.

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Authors and Affiliations

  1. College of Chemistry and Material Science, Shandong Agricultural University, Tai′an, 271018, People’s Republic of China

    Huanshun Yin, Yan Chen, Fei Li & Shiyun Ai

  2. College of Chemistry and Chemical Engineering, Central South University, 410083, Changsha, Hunan, People’s Republic of China

    Hanwen Wu

  3. College of Resources and Environment, Key Laboratory of Agricultural Environment in Universities of Shandong, Shandong Agricultural University, Tai′an, 271018, People’s Republic of China

    Jun Wang

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  1. Huanshun Yin
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Correspondence to Jun Wang.

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Yin, H., Wu, H., Chen, Y. et al. Photoelectrochemical determination of the activity of histone acetyltransferase and inhibitor screening by using MoS2 nanosheets. Microchim Acta 186, 663 (2019). https://doi.org/10.1007/s00604-019-3756-3

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