Developing Novel Fluorescent Probes for Detecting Histone Deacetylase Activity

Project: Research

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Histone Deacetylases (HDAC) are key enzymes in eukaryotic cells that play an essential role in theregulation of gene transcription. HDACs are widely recognized as a major target for the development ofdrugs against a number of diseases, particularly cancer. Numerous reports have shown that abnormalexpression of HDAC is observed in various types of cancers. Due to the important roles of HDACs,many methods have been developed to detect HDAC activity. Among these methods, fluorescence probeshave proven best suited to the task as they offer simple procedures and high sensitivity. Notwithstanding,most of the fluorescence probes for detecting HDAC require an extra step for fluorescent development,thus limiting their real-time application to monitor HDAC activity in cellular environment. Furthermore,there is still a lack of isoform-selective probes for detecting individual HDAC.In this project, a panel of fluorescent probes containing acetyl-lysine and O-NBD group will be firstdesigned and synthesized to detect HDAC activity. Upon incubating the probe with HDAC, deacetylationwill occur and the free amine group will be released. The amine will then react with O-NBD moiety andturn on the fluorescence. The sensitivity and selectivity of the probes will be thoroughly examined.Subsequently the probes will be applied to live cell imaging studies.At the second stage of the proposal, we will develop isoform-selective probes for detecting theactivity of selected HDACs. First we will introduce a selectivity feature by inserting the key probe intoshort peptide sequences that were previously identified as preferred substrate for individual HDAC.Second, we will synthesize a small library of peptide-based probes for screening the substrate specificityof various HDACs. By doing so, we will be able to identify the selective probes for a given HDAC.Based on isoform-selective probes, we will further develop dual function probes by equipping theprobe with an alkyne-containing diazirine photo-crosslinker group. The dual function probes will not onlyenable us to monitor intracellular activity of HDAC, but also allow quantitative characterization of theenzyme’s expression levels in various biological samples. We envision that the probes developed in thisstudy will generate valuable tools for systematic characterization of HDAC in biomedical research. Thecurrent research team has extensive experience in fluorescent probe, peptide chemistry and quantitativeproteomics. We are confident that the research will be completed successfully and provide robust tools tofurther advance HDAC research and facilitate anti-cancer drug development.


Project number9042398
Grant typeGRF
Effective start/end date1/09/164/02/21

    Research areas

  • fluorescent probe , peptide microarray , substrate specificity , histone deacetylase , chemical biology