title: Computational modeling of structurally conserved cancer mutations in the RET and MET kinases: the impact on protein structure, dynamics, and stability.
abstract: Structural and biochemical characterization of protein kinases that confer oncogene addiction and harbor a large number of disease-associated mutations, including RET and MET kinases, have provided insights into molecular mechanisms associated with the protein kinase activation in human cancer. In this article, structural modeling, molecular dynamics, and free energy simulations of a structurally conserved mutational hotspot, shared by M918T in RET and M1250T in MET kinases, are undertaken to quantify the molecular mechanism of activation and the functional role of cancer mutations in altering protein kinase structure, dynamics, and stability. The mechanistic basis of the activating RET and MET cancer mutations may be driven by an appreciable free energy destabilization of the inactive kinase state in the mutational forms. According to our results, the locally enhanced mobility of the cancer mutants and a higher conformational entropy are counterbalanced by a larger enthalpy loss and result in the decreased thermodynamic stability. The computed protein stability differences between the wild-type and cancer kinase mutants are consistent with circular dichroism spectroscopy and differential scanning calorimetry experiments. These results support the molecular mechanism of activation, which causes a detrimental imbalance in the dynamic equilibrium shifted toward the active form of the enzyme. Furthermore, computer simulations of the inhibitor binding with the oncogenic and drug-resistant RET mutations have also provided a plausible molecular rationale for the observed differences in the inhibition profiles, which is consistent with the experimental data. Finally, structural mapping of RET and MET cancer mutations and the computed protein stability changes suggest a similar mechanism of activation, whereby the cancer mutations which display the higher oncogenic activity tend to have the greatest destabilization effect on the inactive kinase structure.
authors.affiliation: Center for Bioinformatics, The University of Kansas, Lawrence, Kansas, USA.
authors.forename: Anshuman
authors.initials: A
authors.lastname: Dixit
authors.forename: Ali
authors.initials: A
authors.lastname: Torkamani
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Schork
authors.forename: Gennady
authors.initials: G
authors.lastname: Verkhivker
mesh_terms: D003198:Computer Simulation; D017124:Conserved Sequence; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D004789:Enzyme Activation / Q000235:genetics; D008958:Models, Molecular; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009369:Neoplasms / Q000201:enzymology* / Q000235:genetics*; D011487:Protein Conformation; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism; D055550:Protein Stability; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019859:Proto-Oncogene Proteins c-met; D051096:Proto-Oncogene Proteins c-ret / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D050505:Mutant Proteins; D047428:Protein Kinase Inhibitors; D011518:Proto-Oncogene Proteins; D017978:Receptors, Growth Factor; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met; D051096:Proto-Oncogene Proteins c-ret