title: In vitro DNA binding studies of antiretroviral drug nelfinavir using ethidium bromide as fluorescence probe.
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.
authors: Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India. Electronic address: ramanammv@yahoo.com.
authors: M M V
authors: MM
authors: Ramana
authors: Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.
authors: Rahul
authors: R
authors: Betkar
authors: Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.
authors: Amey
authors: A
authors: Nimkar
authors: Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.
authors: Prasanna
authors: P
authors: Ranade
authors: Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.
authors: Balaji
authors: B
authors: Mundhe
authors: Department of Chemistry, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098, India.
authors: Sachin
authors: S
authors: Pardeshi
mesh_terms: D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004996:Ethidium / Q000737:chemistry / Q000378:metabolism*; D005399:Fishes / Q000235:genetics; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000378:metabolism; D006860:Hydrogen Bonding; D019888:Nelfinavir / Q000737:chemistry* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D055672:Static Electricity; D013816:Thermodynamics; D014783:Viscosity
chemical_list: D005456:Fluorescent Dyes; D017320:HIV Protease Inhibitors; D004247:DNA; D004996:Ethidium; D019888:Nelfinavir
keywords: Hs DNA; Hydrogen bonding; Minor groove binding; Nelfinavir