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National Institute of Environmental Health Sciences, National Institutes of Health

2 ARTICLES PUBLISHED IN JoVE

Chemistry

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

Akira Sassa¹, William A. Beard¹, David D. Shock¹, Samuel H. Wilson¹

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

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.

Biochemistry

Preparation of Nucleosome Core Particles Complexed with DNA Repair Factors for Cryo-Electron Microscopy Structural Determination

Yesenia Rodriguez¹, Kevin John Butay^*1, Kedar Sharma^*1, Elizabeth Viverette^*1, Samuel H. Wilson¹

¹Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health

This protocol outlines in detail the preparation of nucleosomal complexes using two methods of sample preparation for freezing TEM grids.