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In vitro DNA binding studies of antiretroviral drug nelfinavir using ethidium bromide as fluorescence probe.

作者信息

Ramana M M V, Betkar Rahul, Nimkar Amey, Ranade Prasanna, Mundhe Balaji, Pardeshi Sachin

机构信息

Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.

Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.

出版信息

J Photochem Photobiol B. 2015 Oct;151:194-200. doi: 10.1016/j.jphotobiol.2015.08.012. Epub 2015 Aug 12.

DOI:10.1016/j.jphotobiol.2015.08.012
PMID:26310356
Abstract

Understanding the interaction of small molecules with DNA has become an active research area at the interface between biology and chemistry. In the present work, we investigated the mode of interaction of nelfinavir (NFV) with herring sperm DNA (hs DNA) under physiological conditions using various biophysical techniques. Analysis of UV-absorption and fluorescence spectra indicates the formation of complex between NFV and hs DNA. According to the fluorescence results, the binding constant (K) between NFV and hs DNA was found to be 3.30 × 10(4)LM(-1). The calculated thermodynamic parameters (ΔH° and ΔS°) suggested that hydrogen bonding plays a major role in binding between them. Phosphate group binding studies revealed that there was no electrostatic interactions occurred between NFV and hs DNA. Circular dichroism (CD) and DNA melting curve were employed to measure the conformational change of hs DNA in the presence of NFV, which verified the minor groove binding mode. These results were further supported by viscosity measurements and competitive displacement assay study using Hoechst 33258. According to the sequence specificity experiments, NFV binds to A-T rich region of hs DNA.

摘要

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