Fluorogenic probes for detecting deacylase and demethylase activity towards post-translationally-modified lysine residues |
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Authors: | Yuichiro Hori Miyako Nishiura Tomomi Tao Reisuke Baba Steven D Bull Kazuya Kikuchi |
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Institution: | Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871 Japan.; IFReC, Osaka University, Suita, Osaka 565-0871 Japan ; Department of Chemistry, University of Bath, Bath BA27AY UK ; Quantum Information and Quantum Biology Division, Osaka University, Suita, Osaka 565-0871 Japan |
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Abstract: | Reversible enzymatic post-translational modification of the ε-amino groups of lysine residues (e.g. N-acylation reactions) plays an important role in regulating the cellular activities of numerous proteins. This study describes how enzyme catalyzed N-deprotection of lysine residues of non-fluorescent peptide-coumarin probes can be used to generate N-deprotected peptides that undergo spontaneous O- to N-ester transfer reactions (uncatalyzed) to generate a highly fluorescent N-carbamoyl peptide. This enables detection of enzyme catalyzed N-deacetylation, N-demalonylation, N-desuccinylation and N-demethylation reactions activities towards the N-modified lysine residues of these probes using simple ‘turn on’ fluorescent assays.We developed “turn-on” fluorescent probes that detect enzymatic lysine deacylation and demethylation critical for epigenetic and other cellular phenomena, using intramolecular O- to N-ester transfer reactions. |
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