(627b) Structural, Thermodynamics and Kinetics Role of Novel Hot-Spot Mutations of BCR-ABL1 in Resistance Towards "Ibs" Inhibitors
Specifically, in silicoexperiments will disclose the effects exerted by the âhot-spotâ BCR-ABL1 mutations on the protein structure, thermodynamic stability, and ability to interfere with BCR-ABL1 binding to specific TKIs (e.g., imatinib, dasatinib, and ponatinib).
Structural biology evidences of wild type and mutant isolated KDs and SH2-linker-SH3-KD (SSK) constructs (per seand in complex with TKIs) will be discussed to support in silico predictions.
Direct drug binding to both KD and SSK constructs will be measured using isothermal titration calorimetry and surface plasmon resonance.
In vitrokinase assays will monitor the ability of these mutant isoforms to auto-phosphorylate and to phosphorylate a substrate peptide.
âPrototypicalâ BCR-ABL1 KD mutations will be analyzed in parallel for comparison and to confirm/dispute the current, classical view of TKI resistance.
 Gibbons DL, Pricl S, Posocco P, et al., Molecular dynamics reveal BCR-ABL1 polymutants as a unique mechanism of resistance to PAN-BCR-ABL1 kinase inhibitor therapy. Proc Natl Acad Sci U S A. 2014 Mar 4;111(9):3550-5. doi: 10.1073/pnas.1321173111.