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

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

DOI:
10.3791/50695-v

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14:27 min

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August 19, 2013

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, , ,
1Laboratory of Structural Biology,NIEHS, National Institutes of Health

Capitoli

  • 00:05Titolo
  • 01:55Sample Preparation and Measurement of Steady-state Time Course and Active Site Titration of OGG1
  • 05:53Measurement of Pre-steady-state Time Course
  • 10:56Single-turnover Time Course
  • 11:50Results: Kinetic Measurements of DNA Glycosylase Activity
  • 14:01Conclusion

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.

Tags

DNA GlycosylaseOGG18-oxoGSteady-state KineticsPre-steady-state KineticsSingle-turnover KineticsBurst KineticsProduct ReleaseEnzyme CatalysisDNA Lesion Excision

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