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Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein
 

Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein

Article DOI: 10.3791/1874-v • 11:01 min • March 31st, 2010
March 31st, 2010 •
1, 1, 1, 1
1Molecular Biology and Biochemistry Department, Wesleyan University

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Msh2-Msh6 is responsible for initiating repair of replication errors in DNA. Here we present a transient kinetics approach towards understanding how this critical protein works. The report illustrates stopped-flow experiments for measuring the coupled DNA binding and ATPase kinetics underlying Msh2-Msh6 mechanism of action in DNA repair.

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Stopped-flow Kinetics Methods Mechanism Of Action DNA Repair Protein Transient Kinetic Analysis Biological Macromolecules Active Site Concentrations Intrinsic Rate Constants Macromolecular Function Enzymes Pre-steady State Measurements Reaction Pathway Steady State Measurements Catalytic Efficiency Catalytic Specificity Protein-protein Interactions Protein-ligand Interactions Rate-limiting Conformational Changes Millisecond Timescale Stopped-flow Method Chemical-quench Flow Methods Optical Signal Fluorescence Substrate Binding Product Release Catalytic Turnover Msh2-Msh6 Protein Eukaryotic DNA Repair Protein Base-pair Mismatches Insertion/deletion Loops In DNA Mismatch Repair (MMR) Genomic Integrity
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