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Peptide‐Based Fluorescent Probes for Deacetylase and Decrotonylase Activity: Toward a General Platform for Real‐Time Detection of Lysine Deacylation
Author(s) -
Rooker Debra R.,
Klyubka Yuliya,
Gautam Ritika,
Tomat Elisa,
Buccella Daniela
Publication year - 2018
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700582
Subject(s) - acetylation , sirtuin , lysine , chemistry , peptide , histone deacetylase , biochemistry , histone , nad+ kinase , nicotinamide adenine dinucleotide , cofactor , fluorescence , histone h3 , small molecule , analyte , combinatorial chemistry , histone deacetylase 2 , enzyme , dna , amino acid , physics , quantum mechanics , gene
Histone deacetylases regulate the acetylation levels of numerous proteins and play key roles in physiological processes and disease states. In addition to acetyl groups, deacetylases can remove other acyl modifications on lysines, the roles and regulation of which are far less understood. A peptide‐based fluorescent probe for single‐reagent, real‐time detection of deacetylase activity that can be readily adapted for probing broader lysine deacylation, including decrotonylation, is reported. Following cleavage of the lysine modification, the probe undergoes rapid intramolecular imine formation that results in marked optical changes, thus enabling convenient detection of deacylase activity with good statistical Z′ factors for both absorption and fluorescence modalities. The peptide‐based design offers broader isozyme scope than that of small‐molecule analogues, and is suitable for probing both metal‐ and nicotinamide adenine dinucleotide (NAD + )‐dependent deacetylases. With an effective sirtuin activity assay in hand, it is demonstrated that iron chelation by Sirtinol, a commonly employed sirtuin inhibitor, results in an enhancement in the inhibitory activity of the compound that may affect its performance in vivo.