A homogeneous nonisotopic histone deacetylase activity assay

被引:25
作者
Heltweg, B [1 ]
Jung, M [1 ]
机构
[1] Univ Munster, Dept Pharmaceut & Med Chem, D-48149 Munster, Germany
关键词
histone deacetylase; NDA; fluorescence quenching; homogeneous assay;
D O I
10.1177/1087057102239644
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Historic deacetylases (HDACs) are important regulators of transcription, and their inhibitors are a promising class of anticancer agents. The methods for the determination of HDAC activity and its inhibition that are currently available suffer from various drawbacks, such as animal testing, radioactive substrates, or limited throughput. Therefore, a fast nonisotopic method for the measurement of HDAC activity is highly desirable. The authors present such an assay that relies on the fluorescent HDAC substrate developed previously in their group. After incubation of the substrate with the enzyme, a derivatization leads to efficient fluorescence quenching in the deacetylated metabolite. Thus, only the fluorescence emitted by the remaining substrate is detected, which allows for a convenient detection of HDAC activity in a homogeneous format that can be performed on multiwell plate readers. This procedure, called HDASH (histone deacetylase assay-homogeneous), should be a valuable tool in transcriptional research and especially drug discovery.
引用
收藏
页码:89 / 95
页数:7
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