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

文献类型： 外文期刊

作者： Zhang, Yanmei 1 ; Li, Feng 1 ; Kang, Jingwu 1 ;

作者机构： 1.Chinese Acad Sci, Shanghai Inst Organ Chem, State Key Lab Bioorgan & Nat Prod Chem, Lingling Rd 345, Shanghai 200032, Peoples R China

2.Shanghai Acad Agr Sci, Lab Qual & Safety Risk Assessment Agroprod Shangh, Inst Agri Food Stand & Testing Technol, Minist Agr, Jinqi Rd 1000, Shanghai 201403, Peoples R China

3.ShanghaiTech Univ, Sch Phys Sci, Haike Rd 100, Shanghai 200031, Peoples R China

期刊名称：ANALYTICAL METHODS （ 影响因子：2.896； 五年影响因子：2.716 ）

ISSN： 1759-9660

年卷期： 2017 年 9 卷 37 期

页码：

收录情况： SCI

摘要： 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 degrees 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.

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