È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.
Capitoli
- 00:00Titolo
- 00:57Introduction
- 01:40Sample Preparation for the Msh2-Msh6/ DNA Binding Experiment
- 03:07Instrument Preparation for the Msh-2Msh6/DNA Binding Experiment
- 05:07Msh2-Msh6/DNA Binding Experiment
- 06:27Representative Results for the Msh2-Msh6/DNA Binding Kinietics Experiment
- 07:00Sample and Instrument Preparation for the Msh2-Msh6 ATPase Experiment
- 08:30Msh2-Msh6 ATPase Experiment
- 09:40Representative Results for Msh2-Msh6 ATPase Kinetics Experiments
- 10:20Conclusion
MSH2-MSH6 è responsabile per l'avvio di riparazione di errori di replicazione del DNA. Qui vi presentiamo un approccio transitorio cinetica verso la comprensione di come lavora questa proteina critica. Il rapporto illustra stopped-flow esperimenti per misurare il legame del DNA accoppiati e cinetica ATPasi sottostante MSH2-MSH6 meccanismo d'azione nella riparazione del DNA.
Tags
Stopped-flow Kinetics MethodsMechanism Of ActionDNA Repair ProteinTransient Kinetic AnalysisBiological MacromoleculesActive Site ConcentrationsIntrinsic Rate ConstantsMacromolecular FunctionEnzymesPre-steady State MeasurementsReaction PathwaySteady State MeasurementsCatalytic EfficiencyCatalytic SpecificityProtein-protein InteractionsProtein-ligand InteractionsRate-limiting Conformational ChangesMillisecond TimescaleStopped-flow MethodChemical-quench Flow MethodsOptical SignalFluorescenceSubstrate BindingProduct ReleaseCatalytic TurnoverMsh2-Msh6 ProteinEukaryotic DNA Repair ProteinBase-pair MismatchesInsertion/deletion Loops In DNAMismatch Repair (MMR)Genomic Integrity
