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Issue 37, 2017
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Screening of histone deacetylase 1 inhibitors in natural products by capillary electrophoresis

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Abstract

A method for the screening of histone deacetylase 1 (HDAC1) inhibitors in natural products by using capillary electrophoresis (CE) coupled with laser induced fluorescence (LIF) detection was developed. The method was developed by employing a 5-carboxyfluorescein labelled peptide with an acetylated lysine residue as the substrate of HDAC1 and a small chemical library composed of 38 purified natural products. The biochemical assay was performed by means of CE separation, i.e. the deacetylated product in the enzymatic reaction solution was separated from the substrate peptide; therefore, the enzyme activity can be accurately calculated through the measurement of the peak area of the deacetylated product. For inhibitor screening, the tested samples were spiked in the substrate solution and the resulting solution mixtures were then incubated with the enzyme solution for proceeding the deacetylation reaction for a short period of time at 30 °C. After quenching the reaction by putting the reaction vial in boiling water, the resulting solution was injected for CE separation with 100 mM Tris–H3PO4 buffer (pH 8.0) containing 30 mM NaCl and 0.05% (m/v) hexadimethrine bromide (HDB). The inhibitor could be readily identified as long as the peak area of the deacetylated product is reduced in comparison with that of the negative control in the absence of any inhibitor. Dynamically coating the capillary wall with positively charged HDB is very necessary to diminish the serious adsorption of the substrate and product peptides onto the capillary wall. A rapid, cost-effective method for HDAC1 inhibitor screening is proposed.

Graphical abstract: Screening of histone deacetylase 1 inhibitors in natural products by capillary electrophoresis

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Publication details

The article was received on 08 Jun 2017, accepted on 19 Aug 2017 and first published on 21 Aug 2017


Article type: Paper
DOI: 10.1039/C7AY01428G
Citation: Anal. Methods, 2017,9, 5502-5508
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    Screening of histone deacetylase 1 inhibitors in natural products by capillary electrophoresis

    Y. Zhang, F. Li and J. Kang, Anal. Methods, 2017, 9, 5502
    DOI: 10.1039/C7AY01428G

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