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- Title
A Qualitative and Quantitative Assay to Study DNA/Drug Interaction Based on Sequence Selective Inhibition of Restriction Endonucleases.
- Authors
Hassan, Syed A.; Chauhan, Lata; Barthwal, Ritu; Dixit, Aparna
- Abstract
Purpose: To explore the use of restriction inhibition assay (RIA) to study the binding specificity of some anticancer drugs. Methods: A 448 bp DNA fragment derived from pBCKS+ plasmid (harboring the polylinker region with multiple restriction endonuclease sites) was used as a template for sequence selective inhibition of the test drugs. The template was incubated with different concentrations of anticancer drugs (adriamycin, daunomycin, mitoxantrone, distamycin-A, berberine and palmatine) prior to digestion with restriction endonucleases - HindIII, EcoRI and EcoRV. Results: Mitoxantrone, adriamycin and daunomycin showed specificity for HindIII restriction site (5'- AAGCTT-3') at 220, 100 and 100 µM concentration, respectively. Conversely, distamycin-A showed an affinity for EcoRI (5'-AAATGC-3') restriction sites at a concentration of 10 µM. No binding was observed for berberine and palmatine at a maximum concentration of 2 mM at HindIII, EcoRI and EcoRV restriction sites, respectively. Conclusion: The inhibition of endonucleases by mitoxantrone, adriamycin, daunomycin, distamycin-A, provides direct evidence of the co-existence of concentration and sequence specificity for drug-DNA interaction as well as the need to explore the possible use of RIA for demonstrating the binding specificity of anticancer drugs.
- Subjects
NUCLEOTIDE sequence; DRUG interactions; ANTINEOPLASTIC agents; ENDONUCLEASE genetics; MITOXANTRONE; DAUNOMYCIN; BINDING sites
- Publication
Tropical Journal of Pharmaceutical Research, 2012, Vol 11, Issue 5, p721
- ISSN
1596-5996
- Publication type
Article
- DOI
10.4314/tjpr.v11i5.4