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Molecules 2015, 20(1), 1176-1191; doi:10.3390/molecules20011176

Kinase Inhibitor Profile for Human Nek1, Nek6, and Nek7 and Analysis of the Structural Basis for Inhibitor Specificity

1
LaboratórioNacional de Biociências, Centro Nacional de PesquisaemEnergia e Materiais, Campinas, 13083-970 SP, Brazil
2
Programa de Pós-graduação em Biologia Funcional e Molecular, Departamento de Bioquímica e BiologiaTecidual, Instituto de Biologia, UniversidadeEstadual de Campinas, Campinas, 13083-862 SP, Brazil
3
Instituto Carlos Chagas, FIOCRUZ, Curitiba, 81350-010 PR, Brazil
4
Departamento de Bioquímica e de Biologia Tecidual, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, 13083-862 SP, Brazil
5
Faculdade de Ciências Farmacêuticas, Universidade Estadual de Campinas, Campinas, 13083-859 SP, Brazil
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Jean Jacques Vanden Eynde
Received: 4 November 2014 / Accepted: 5 January 2015 / Published: 13 January 2015
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Abstract

Human Neks are a conserved protein kinase family related to cell cycle progression and cell division and are considered potential drug targets for the treatment of cancer and other pathologies. We screened the activation loop mutant kinases hNek1 and hNek2, wild-type hNek7, and five hNek6 variants in different activation/phosphorylation statesand compared them against 85 compounds using thermal shift denaturation. We identified three compounds with significant Tm shifts: JNK Inhibitor II for hNek1(Δ262-1258)-(T162A), Isogranulatimide for hNek6(S206A), andGSK-3 Inhibitor XIII for hNek7wt. Each one of these compounds was also validated by reducing the kinases activity by at least 25%. The binding sites for these compounds were identified by in silico docking at the ATP-binding site of the respective hNeks. Potential inhibitors were first screened by thermal shift assays, had their efficiency tested by a kinase assay, and were finally analyzed by molecular docking. Our findings corroborate the idea of ATP-competitive inhibition for hNek1 and hNek6 and suggest a novel non-competitive inhibition for hNek7 in regard to GSK-3 Inhibitor XIII. Our results demonstrate that our approach is useful for finding promising general and specific hNekscandidate inhibitors, which may also function as scaffolds to design more potent and selective inhibitors. View Full-Text
Keywords: drug discovery; compound screening; kinase assays; molecular modeling; molecular docking; ATP-competitive inhibitors; non-competitive inhibition; Neks drug discovery; compound screening; kinase assays; molecular modeling; molecular docking; ATP-competitive inhibitors; non-competitive inhibition; Neks
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MDPI and ACS Style

Moraes, E.C.; Meirelles, G.V.; Honorato, R.V.; de Souza, T.A.C.B.; de Souza, E.E.; Murakami, M.T.; de Oliveira, P.S.L.; Kobarg, J. Kinase Inhibitor Profile for Human Nek1, Nek6, and Nek7 and Analysis of the Structural Basis for Inhibitor Specificity. Molecules 2015, 20, 1176-1191.

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