Cysteine Modification by Ebselen Reduces the Stability and Cellular Levels of 14-3-3 Proteins

被引:8
作者
Waloen, Kai [1 ]
Jung-KC, Kunwar [1 ]
Vecchia, Elisa D. [3 ]
Pandey, Sunil [1 ]
Gasparik, Norbert [4 ]
Doskeland, Anne [1 ,2 ]
Patil, Sudarshan [1 ]
Kleppe, Rune [5 ,6 ]
Hritz, Jozef [4 ]
Norton, William H. J. [3 ]
Martinez, Aurora [1 ]
Haavik, Jan [1 ,7 ]
机构
[1] Univ Bergen, Dept Biomed, N-5009 Bergen, Norway
[2] Univ Bergen, Prote Unit PROBE, Bergen, Norway
[3] Univ Leicester, Coll Med Biol Sci & Psychol, Dept Neurosci Psychol & Behav, Leicester, Leics, England
[4] Masaryk Univ, CEITEC MU, Brno, Czech Republic
[5] Masaryk Univ, Fac Sci, Dept Chem, Brno, Czech Republic
[6] Haukeland Hosp, Norwegian Ctr Maritime & Diving Med, Dept Occupat Med, Bergen, Norway
[7] Haukeland Hosp, Div Psychiat, Bergen, Norway
基金
欧盟地平线“2020”;
关键词
TYROSINE-HYDROXYLASE; STRUCTURAL BASIS; PHOSPHORYLATION; 14-3-3-PROTEINS; IDENTIFICATION; PROLIFERATION; THIMEROSAL; INHIBITOR; OXIDATION; TARGETS;
D O I
10.1124/molpharm.120.000184
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
The 14-3-3 proteins constitute a family of adaptor proteins with many binding partners and biological functions, and they are considered promising drug targets in cancer and neuropsychiatry. By screening 1280 small-molecule drugs using differential scanning fluorimetry (DSF), we found 15 compounds that decreased the thermal stability of 14-3-3(. Among these compounds, ebselen was identified as a covalent, destabilizing ligand of 14-3-3 isoforms (, e, y, and n. Ebselen bonding decreased 14-3-3( binding to its partner Ser19-phosphorylated tyrosine hydroxylase. Characterization of site-directed mutants at cysteine residues in 14-3-3( (C25, C94, and C189) by DSF and mass spectroscopy revealed covalent modification by ebselen of all cysteines through a selenylsulfide bond. C25 appeared to be the preferential site of ebselen interaction in vitro, whereas modification of C94 was the main determinant for protein destabilization. At therapeutically relevant concentrations, ebselen and ebselen oxide caused decreased 14-3-3 levels in SH-SY5Y cells, accompanied with an increased degradation, most probably by the ubiquitin-dependent proteasome pathway. Moreover, ebselen-treated zebrafish displayed decreased brain 14-3-3 content, a freezing phenotype, and reduced mobility, resembling the effects of lithium, consistent with its proposed action as a safer lithium-mimetic drug. Ebselen has recently emerged as a promising drug candidate in several medical areas, such as cancer, neuropsychiatric disorders, and infectious diseases, including coronavirus disease 2019. Its pleiotropic actions are attributed to antioxidant effects and formation of selenosulfides with critical cysteine residues in proteins. Our work indicates that a destabilization of 14-3-3 may affect the protein interaction networks of this protein family, contributing to the therapeutic potential of ebselen.
引用
收藏
页码:155 / 169
页数:15
相关论文
共 65 条
[1]   14-3-3 proteins: A historic overview [J].
Aitken, Alastair .
SEMINARS IN CANCER BIOLOGY, 2006, 16 (03) :162-172
[2]   Discovery of a Specific Inhibitor of Pyomelanin Synthesis in Legionella pneumophila [J].
Aubi, Oscar ;
Flydal, Marte I. ;
Zheng, Huaixin ;
Skjaerven, Lars ;
Rekand, Illimar ;
Leiros, Hanna-Kirsti S. ;
Haug, Bengt Erik ;
Cianciotto, Nicholas P. ;
Martinez, Aurora ;
Underhaug, Jarl .
JOURNAL OF MEDICINAL CHEMISTRY, 2015, 58 (21) :8402-8412
[3]   Ebselen, a promising antioxidant drug: mechanisms of action and targets of biological pathways [J].
Azad, Gajendra Kumar ;
Tomar, Raghuvir S. .
MOLECULAR BIOLOGY REPORTS, 2014, 41 (08) :4865-4879
[4]   14-3-3: A Case Study in PPI Modulation [J].
Ballone, Alice ;
Centorrino, Federica ;
Ottmann, Christian .
MOLECULES, 2018, 23 (06)
[5]   Small-Molecule Stabilization of the 14-3-3/Gab2 Protein-Protein Interaction (PPI) Interface [J].
Bier, David ;
Bartel, Maria ;
Sies, Katharina ;
Halbach, Sebastian ;
Higuchi, Yusuke ;
Haranosono, Yu ;
Brummer, Tilman ;
Kato, Nobuo ;
Ottmann, Christian .
CHEMMEDCHEM, 2016, 11 (08) :911-918
[6]   QUANTITATIVE-ANALYSIS OF PROTEIN FAR UV CIRCULAR-DICHROISM SPECTRA BY NEURAL NETWORKS [J].
BOHM, G ;
MUHR, R ;
JAENICKE, R .
PROTEIN ENGINEERING, 1992, 5 (03) :191-195
[7]   Down-regulation of 14-3-3β exerts anti-cancer effects through inducing ER stress in human glioma U87 cells: Involvement of CHOP-Wnt pathway [J].
Cao, Lei ;
Lei, Hui ;
Chang, Ming-Ze ;
Liu, Zhi-Qin ;
Bie, Xiao-Hua .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2015, 462 (04) :389-395
[8]   The cysteine-reactive small molecule ebselen facilitates effective SOD1 maturation [J].
Capper, Michael J. ;
Wright, Gareth S. A. ;
Barbieri, Letizia ;
Luchinat, Enrico ;
Mercatelli, Eleonora ;
McAlary, Luke ;
Yerbury, Justin J. ;
O'Neill, Paul M. ;
Antonyuk, Svetlana V. ;
Banci, Lucia ;
Hasnain, S. Samar .
NATURE COMMUNICATIONS, 2018, 9
[9]   p53 suppresses 14-3-3γ by stimulating proteasome-mediated 14-3-3γ protein degradation [J].
Chen, De-Yu ;
Dai, Dong-Fang ;
Hua, Ye ;
Qi, Wen-Qing .
INTERNATIONAL JOURNAL OF ONCOLOGY, 2015, 46 (02) :818-824
[10]   14-3-3 Proteins in Brain Development: Neurogenesis, Neuronal Migration and Neuromorphogenesis [J].
Cornell, Brett ;
Toyo-Oka, Kazuhito .
FRONTIERS IN MOLECULAR NEUROSCIENCE, 2017, 10