Myeloid cell leukemia-1 (Mcl-1) is an anti-apoptotic member of the Bcl-2 family proteins. Its amplification is one of the most frequent genetic aberrations found in human cancers. Pyridoclax, a promising BH3 mimetic inhibitor, interacts directly with Mcl-1 and induces massive apoptosis at a concentration of 15 µM in combination with anti-Bcl-xL strategies in chemo-resistant ovarian cancer cell lines. In this study, a combined experimental and theoretical approach was used to investigate the binding mode of Pyridoclax to Mcl-1. The representative poses generated from dynamics simulations compared with NMR data revealed: (i) Pyridoclax bound to P1 and P2 pockets of Mcl-1 BH3 binding groove through its styryl a... More
Myeloid cell leukemia-1 (Mcl-1) is an anti-apoptotic member of the Bcl-2 family proteins. Its amplification is one of the most frequent genetic aberrations found in human cancers. Pyridoclax, a promising BH3 mimetic inhibitor, interacts directly with Mcl-1 and induces massive apoptosis at a concentration of 15 µM in combination with anti-Bcl-xL strategies in chemo-resistant ovarian cancer cell lines. In this study, a combined experimental and theoretical approach was used to investigate the binding mode of Pyridoclax to Mcl-1. The representative poses generated from dynamics simulations compared with NMR data revealed: (i) Pyridoclax bound to P1 and P2 pockets of Mcl-1 BH3 binding groove through its styryl and methyl groups establishing mainly hydrophobic contacts, (ii) one of the ending pyridines interacts through electrostatic interaction with K234 side chain, a negatively charged residue present only in this position in Mcl-1.AbbreviationsATCCAmerican Type Culture CollectionBcl-2B-cell lymphoma 2Bcl-xLB-cell lymphoma-extra largeBHBcl-2 HomologyCCPNMRCollaborative Computational Project for NMRCCSPCombined Chemical Shift PerturbationsCGENFFCHARMM GENeral Force FieldCHARMMChemistry At Harvard Molecular MechanicsDAPI4',6'-diamidino-2-phenylindoleDMSO-d6Deuterated DimethylsulfoxideDTTDithiothreitolEDTAEthylene-Diamine-Tetra-Acetic acidGOLDGenetic Optimization for Ligand DockingHEPES4-(2-HydroxyEthyl)-1-PiperazineEthaneSulfonic acidHSQCHeteronuclear Single Quantum CorrelationIMACImmobilized Metal Ion Affinity ChromatographyIPTGIsopropyl-Beta-D-ThiogalactosideKDDissociation ConstantLPLinear PredictionMBPMaltose-Binding ProteinMcl-1Myeloid cell leukemia-1NMRNuclear Magnetic ResonanceNOENuclear Overhauser EffectNPTNumber, Pressure, and TemperatureNVTNumber, Volume, and TemperaturePARPPoly- Adenosine-DiPhosphate-Ribose-PolymérasePBSPhosphate-Buffer SalinePDBProtein Data BankPMEParticle Mesh EwaldPMSFPhenylMethaneSulfonyl FluoridePVDFPolyVinylidene DiFluorideRMSDRoot-Mean-Square DeviationRPMIRoswell Park Memorial InstituteRTRoom TemperatureSDS-PAGESodium Dodecyl Sulfate Polyacrylamide Gel ElectrophoresisSOFASTband-Selective Optimized Flip Angle Short TransientHMQCHeteronuclear Multiple Quantum CoherenceTBSTris Buffered SalineTITANTITration ANalysisTPPITime Proportional Phase IncrementationCommunicated by Ramaswamy H. Sarma.
