Abstract
Extracellular signal-regulated kinase 8 (ERK8) has been already implicated in cell transformation and in the protection of genomic integrity and, therefore, proposed as a novel potential therapeutic target for cancer. In the absence of a crystal structure, we developed a three-dimensional model for its kinase domain. To validate our model we applied a structure-based virtual screening protocol consisting of pharmacophore screening and molecular docking. Experimental characterization of the hit compounds confirmed that a high percentage of the identified scaffolds was able to inhibit ERK8. We also confirmed an ATP competitive mechanism of action for the two best-performing molecules. Ultimately, we identified an ERK8 drug-resistant "gatekeeper" mutant that corroborated the predicted molecular binding mode, confirming the reliability of the generated structure. We expect that our model will be a valuable tool for the development of specific ERK8 kinase inhibitors.
Publication types
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Research Support, Non-U.S. Gov't
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Validation Study
MeSH terms
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Adenosine Diphosphate / chemistry
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Adenosine Diphosphate / metabolism
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Adenosine Triphosphate / chemistry
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Adenosine Triphosphate / metabolism
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Amino Acid Sequence
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Binding Sites / genetics
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Biocatalysis / drug effects
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Blotting, Western
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Extracellular Signal-Regulated MAP Kinases / chemistry*
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Extracellular Signal-Regulated MAP Kinases / genetics
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Extracellular Signal-Regulated MAP Kinases / metabolism*
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Humans
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Models, Molecular*
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Molecular Dynamics Simulation
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Molecular Sequence Data
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Molecular Structure
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Mutation
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Protein Binding
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Protein Kinase Inhibitors / chemistry
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Protein Kinase Inhibitors / metabolism
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Protein Kinase Inhibitors / pharmacology
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Protein Structure, Tertiary*
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Sequence Homology, Amino Acid
Substances
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Protein Kinase Inhibitors
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Adenosine Diphosphate
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Adenosine Triphosphate
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Extracellular Signal-Regulated MAP Kinases
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MAPK15 protein, human
Grants and funding
This work was supported by a Start-up grant from Regione Toscana and by a grant to Regione Toscana from Ministero della Salute, in the context of the “Programma per la Ricerca Sanitaria 2008: Bando Cellule Staminali”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.