0519605 - BTÚ 2020 RIV US eng J - Článek v odborném periodiku
Kutil, Zsófia - Mikesova, Jana - Zessin, M. - Meleshin, M. - Nováková, Zora - Alquicer, Glenda - Kozikowski, A. - Sippl, W. - Bařinka, Cyril - Schutkowski, M.
Continuous Activity Assay for HDAC11 Enabling Reevaluation of HDAC Inhibitors.
ACS Omega. Roč. 4, č. 22 (2019), s. 19895-19904. ISSN 2470-1343. E-ISSN 2470-1343
Grant CEP: GA ČR GA15-19640S; GA MŠMT(CZ) ED1.1.00/02.0109
Institucionální podpora: RVO:86652036
Klíčová slova: histone deacetylase inhibitors * sir2 family * POSTTRANSLATIONAL MODIFICATIONS
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 2.870, rok: 2019 ; AIS: 0.573, rok: 2019
Způsob publikování: Open access
Web výsledku:
https://pubs.acs.org/doi/10.1021/acsomega.9b02808DOI: https://doi.org/10.1021/acsomega.9b02808

Histone deacetylase 11 (HDAC11) preferentially removes fatty acid residues from lysine side chains in a peptide or protein environment. Here, we report the development and validation of a continuous fluorescence-based activity assay using an internally quenched TNF alpha-derived peptide derivative as a substrate. The threonine residue in the +1 position was replaced by the quencher amino acid 3'-nitro-L-tyrosine and the fatty acyl moiety substituted by 2-aminobenzoylated 11-aminoundecanoic acid. The resulting peptide substrate enables fluorescence-based direct and continuous readout of HDAC11-mediated amide bond cleavage fully compatible with high-throughput screening formats. The Z'-factor is higher than 0.85 for the 15 mu M substrate concentration, and the signal-to-noise ratio exceeds 150 for 384-well plates. In the absence of NAD(+), this substrate is specific for HDAC 11. Reevaluation of inhibitory data using our novel assay revealed limited potency and selectivity of known HDAC inhibitors, including Elevenostat, a putative HDAC11-specific inhibitor.
Trvalý link: http://hdl.handle.net/11104/0305194