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Fluorescence assay of polyamide–DNA interactions
- Source :
-
Analytical Biochemistry . Apr2012, Vol. 423 Issue 1, p178-183. 6p. - Publication Year :
- 2012
-
Abstract
- Abstract: Polyamides (PAs) are distamycin-type ligands of DNA that bind the minor groove and are capable of sequence selective recognition. This capability provides a viable route to their development as therapeutics. Presented here is a simple and convenient fluorescence assay for PA–DNA binding. PAs are titrated into a sample of a hairpin DNA featuring a TAMRA dye attached to an internal dU near the PA binding site. In a study of 6 PAs, PA binding leads to a steady reproducible decrease in fluorescence intensity that can be used to generate binding isotherms. The assay works equally well with both short (6- to 8-ring) and long (14-ring) PAs, and K d values ranging from approximately 1nM to at least 140nM were readily obtained using a simple monochromator or filter configuration. Competition assays provide a means to assessing possible dye interference, which can be negligible. The assay can also be used to determine PA extinction coefficients and to measure binding kinetics; thus, it is an accessible and versatile tool for the study of PA properties and PA–DNA interactions. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 00032697
- Volume :
- 423
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Analytical Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 73491120
- Full Text :
- https://doi.org/10.1016/j.ab.2012.01.017