Abstract
In addition to binding affinity, the on and off rates of drug-DNA interactions are important in determining the biological activities of a drug For example, the rate of dissociation of a drug from DNA has been shown to be related to its pharmacological activities (1). Various techniques have been employed to study the dissociation kinetics of drugs from DNA. These include detergent sequestration technique pioneered by Muller and Crothers (1), a modification of the footprinting technique for examming the dissociation of ligands from individual binding sites (2), relaxation methods such as T-Jump for measuring fast kinetics (3), and a procedure that can yield drug—DNA dissociation kinetics under conditions of active transcription of the DNA (4). The simplest and most widely used method IS detergent-induced dissociation rate measurement incorporating the detergent sodium dodecyl sulfate (SDS). This chapter will thus focus on the SDS-sequestration technique and include only conventional spectropliotometric and stopped-flow methods. An example of an actinomycin D dissociation measurement from the author’s laboratory will be used to illustrate the methodology
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References
Muller W. and Crothers D. M. (1968) Studies of the Binding of Actmomycin and Related Compounds to DNA. J Mol Biol 35, 25l–290.
Fletcher M. C. and Fox K. R. (1993) Visuahsing the Kinetics of Dissociation of Actmomycin from Individual Sites in Mixed Sequence DNA by Dnase I Footprinting Nuclic Acids Res 21, 1339–1344
Chaires J. B., Dattagupta N., and Crothers D. M. (1985) Kinetics of the Daunomycin-DNA Interaction. Biochemistry 24, 260–267.
Phillips D. R. and Crothers D. M. (1986) Kanetics and Sequence Specificity of Drug-DNA Interactions An in Vitro Transcription Assay Biochemistry 25, 7355–7362.
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© 1997 Humana Press Inc., Totowa, NJ
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Chen, FM. (1997). Methods for the Studies of Drug Dissociation from DNA. In: Fox, K.R. (eds) Drug-DNA Interaction Protocols. Methods in Molecular Biology™, vol 90. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-447-X:269
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DOI: https://doi.org/10.1385/0-89603-447-X:269
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-447-1
Online ISBN: 978-1-59259-574-7
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