title: Structure prediction and validation of the ERK8 kinase domain.
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.
authors: Istituto Toscano Tumori-Core Research Laboratory, Signal Transduction Unit, AOU Senese, Siena, Italy.
authors: Angela
authors: A
authors: Strambi
authors: Mattia
authors: M
authors: Mori
authors: Matteo
authors: M
authors: Rossi
authors: David
authors: D
authors: Colecchia
authors: Fabrizio
authors: F
authors: Manetti
authors: Francesca
authors: F
authors: Carlomagno
authors: Maurizio
authors: M
authors: Botta
authors: Mario
authors: M
authors: Chiariello
mesh_terms: D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D055162:Biocatalysis / Q000187:drug effects; D015153:Blotting, Western; D048049:Extracellular Signal-Regulated MAP Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009154:Mutation; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid
chemical_list: D047428:Protein Kinase Inhibitors; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D048049:Extracellular Signal-Regulated MAP Kinases; C456320:MAPK15 protein, human