Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

A Qualitative and Quantitative Assay to Study DNA/Drug Interaction Based on Sequence Selective Inhibition of Restriction Endonucleases

Syed A Hassan¹ , Lata Chauhan², Ritu Barthwal², Aparna Dixit³

¹Faculty of Computing and Information Technology, King Abdul Aziz University, Rabigh-21911, Saudi Arabia; ²Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee-247667, Uttaranchal; ³School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India.

For correspondence:- Syed Hassan Email: asifsrmcbt@yahoo.com Tel:+91544894526

Received: 29 December 2011 Accepted: 21 August 2012 Published: 18 October 2012

Citation: Hassan SA, Chauhan L, Barthwal R, Dixit A. A Qualitative and Quantitative Assay to Study DNA/Drug Interaction Based on Sequence Selective Inhibition of Restriction Endonucleases. Trop J Pharm Res 2012; 11(5):721-727 doi: 10.4314/tjpr.v11i5.4

© 2012 The authors.
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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.

Keywords: Restriction endonucleases, Restriction sites, Anticancer drugs, Restriction inhibition assay (RIA), Binding specificity