Jumonji domain-containing proteins (JMJDs) play an important role in the epigenetic regulation of gene expression. Aberrant regulation of histone modification has been observed in the progression of a variety of diseases, such as neurological disorders and cancer. Therefore, discovery of selective modulators of JMJDs is very attractive in new drug discovery. Herein, a simple capillary electrophoresis (CE) method was developed for screening of inhibitors against JMJD3. A known JMJD3 inhibitor GSK-J1, 5-carboxyfluorescein labeled substrate peptide with an amino acid sequence of KAPRKQLATKAARK(me3)SAPATGG (truncated from histone H3), as well as a small chemical library composed of 37 purified natural compounds and 30 natural extracts were used for method development and validation. The separation of substrate from its demethylated product was achieved by addition of polycation hexadimethrine bromide (HDB) in the running buffer. The enzyme activity was thus assayed accurately through separating the demethylated product from the substrate and then measuring the peak area of the product. The enzyme inhibition can be read out by comparing the peak area of the demethylated product obtained in the present of inhibitors and that of the negative control in the absence of any inhibitor. The merit of the method is proved by discovering two new JMJD3 inhibitors: salvianic acid A and puerarin 6′’-O-xyloside.
公司名称 | 产品信息 | 采购帮参考价格 | |
---|---|---|---|
阿拉丁 | Dimethyl sulfoxide |
|
¥20.00~¥221931.13 |
阿拉丁 | sodium hydroxide |
|
¥15.00~¥23974.27 |
阿拉丁 | sodium fluorescein |
|
¥27.00~¥20650.07 |
Sigma | ascorbic acid |
|
¥15.00~¥20070.64 |
Sigma | triton X-100 |
|
¥20.00~¥9871.90 |
Sigma | Hexadimethrine bromide |
|
¥140.00~¥6742.36 |
阿拉丁 | sodium dihydrogen phosphate |
|
¥72.00~¥308.00 |
上海源叶 | 绿原酸 |
B20782
|
|
上海源叶 | 芦丁 |
|
|
上海源叶 | 槲皮素 |
B20527
|
|
上海源叶 | 没食子酸 |
B20851
|
|
上海源叶 | 迷迭香酸 |
B20862
|
|
上海源叶 | 丹酚酸B |
B20261
|
|
上海源叶 | All 37 purified natural products |
|
|
阿拉丁 | hydroxyethyl piperazine ethyl sulfonic acid |
|
|
Sigma | (NH?)?Fe(SO?)??6H?O |
|