ac501881s_si_002.pdf (965.82 kB)
Development of a Fluorogenic Probe Based on a DNA Staining Dye for Continuous Monitoring of the Histone Deacetylase Reaction
journal contribution
posted on 2014-08-05, 00:00 authored by Masafumi Minoshima, Tetsuaki Matsumoto, Kazuya KikuchiWe
designed a simple, rapid, and continuous method for the detection
of the activity of histone deacetylases (HDACs), which are key enzymes
involved in epigenetic gene regulation, using a DNA-based fluorogenic
probe. We designed and synthesized a fluorogenic probe, BOXTO-GK(Ac)G,
which is a DNA staining dye–peptide conjugate containing an
acetylated lysine. The DNA-dependent fluorescence of BOXTO-GK(Ac)G
was greatly enhanced upon deacetylation of the acetylated lysine moiety,
owing to the increased DNA binding ability of the probe. The HDAC
reaction was detected through a simple procedure that combined this
probe with DNA. Our detection system monitored the enzymatic reaction
in real time and could be applied to the inhibition assay. These findings
demonstrated that our system might be a useful tool for the analysis
of HDAC function and for the evaluation of the inhibitor potencies
of drug candidates that target HDACs.