Nuclear factor (erythroid-derived 2)-like 2 (NRF2) drug discovery: Biochemical toolbox to develop NRF2 activators by reversible binding of Kelch-like ECH-associated protein 1 (KEAP1).

Autor: Bresciani A; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Missineo A; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Gallo M; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Cerretani M; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Fezzardi P; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Tomei L; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Cicero DO; IRBM Science Park S.p.A., Pomezia, Roma, Italy; Dipartimento di Scienze e Tecnologie Chimiche, Università di Roma 'Tor Vergata', Roma, Italy., Altamura S; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Santoprete A; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Ingenito R; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Bianchi E; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Pacifici R; CHDI Management/CHDI Foundation, Los Angeles, CA, USA., Dominguez C; CHDI Management/CHDI Foundation, Los Angeles, CA, USA., Munoz-Sanjuan I; CHDI Management/CHDI Foundation, Los Angeles, CA, USA., Harper S; IRBM Science Park S.p.A., Pomezia, Roma, Italy., Toledo-Sherman L; CHDI Management/CHDI Foundation, Los Angeles, CA, USA., Park LC; CHDI Management/CHDI Foundation, Los Angeles, CA, USA. Electronic address: larry.park@chdifoundation.org.
Jazyk: angličtina
Zdroj: Archives of biochemistry and biophysics [Arch Biochem Biophys] 2017 Oct 01; Vol. 631, pp. 31-41. Date of Electronic Publication: 2017 Aug 08.
DOI: 10.1016/j.abb.2017.08.003
Abstrakt: Mechanisms that activate innate antioxidant responses, as a way to mitigate oxidative stress at the site of action, hold much therapeutic potential in diseases, such as Parkinson's disease, Alzheimer's disease and Huntington's disease, where the use of antioxidants as monotherapy has not yielded positive results. The nuclear factor NRF2 is a transcription factor whose activity upregulates the expression of cell detoxifying enzymes in response to oxidative stress. NRF2 levels are modulated by KEAP1, a sensor of oxidative stress. KEAP1 binds NRF2 and facilitates its ubiquitination and subsequent degradation. Recently, compounds that reversibly disrupt the NRF2-KEAP1 interaction have been described, opening the field to a new era of safer NRF2 activators. This paper describes a set of new, robust and informative biochemical assays that enable the selection and optimization of non-covalent KEAP1 binders. These include a time-resolved fluorescence resonance energy transfer (TR-FRET) primary assay with high modularity and robustness, a surface plasmon resonance (SPR) based KEAP1 direct binding assay that enables the quantification and analysis of full kinetic binding parameters and finally a 1 H- 15 N heteronuclear single quantum coherence (HSQC) NMR assay suited to study the interaction surface of KEAP1 with residue-specific information to validate the interaction of ligands in the KEAP1 binding site.
(Copyright © 2017 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: