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Steady-state, Pre-steady-state, and Single-turnover Kinetic Measurement for DNA Glycosylase Activity
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Steady-state, Pre-steady-state, and Single-turnover Kinetic Measurement for DNA Glycosylase Activity

Article DOI: 10.3791/50695-v • 14:27 min • August 19th, 2013
August 19th, 2013 •
1, 1, 1, 1
1Laboratory of Structural Biology, NIEHS, National Institutes of Health

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Summary
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Time courses for the glycosylase activity of 8-oxoguanine DNA glycosylase are biphasic exhibiting a burst of product formation and a linear steady-state phase. Utilizing quench-flow techniques, the burst and the steady-state rates can be measured, which correspond to excision of 8-oxoguanine and release of the glycosylase from the product DNA, respectively.

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Steady-state Pre-steady-state Single-turnover Kinetic Measurement DNA Glycosylase Activity OGG1 8-oxoguanine 8-oxoG Product Formation Burst Phase Steady-state Phase Enzyme Concentration Intrinsic Rate Product Release Rate DNA Dissociation Rate Constant Koff Catalytic Cycling Fluorescent Labeled Lesion-containing Oligonucleotide Purified OGG1 Kinetic Measurements
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