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Binding of protoberberine alkaloid coralyne with double stranded poly(A): a biophysical study

Content Provider	Royal Society of Chemistry (RSC)
Author	Giri, Prabal Kumar, Gopinatha Suresh
Copyright Year	2008
Abstract	Recognition of double stranded ribonucleic acid is a critical event in many biological pathways such as trafficking, editing and maturation of mRNA, interferon antiviral response and RNA interference. In the context of probing double stranded RNA binding small molecules, the interaction of the antitumor protoberberine alkaloidcoralyne with double stranded poly(A) has been studied by various biophysical techniques. Typical hypochromic and bathochromic shifts in the absorption spectrum and appreciable quenching of the intrinsic fluorescence of coralyne indicated the strong affinity of coralyne to poly(A). The corresponding intrinsic binding constant evaluated from Scatchard analysis was in the order of 105 M−1. The strong binding was further characterized by significant polarization of the alkaloid fluorescence and stabilization of poly(A) helix against thermal strand separation. The binding process was manifested by remarkable perturbation of the intrinsic circular dichroic spectrum of poly(A) with concomitant generation of optical activity in the bound alkaloid molecules that are otherwise achiral. Job plot analysis showed the binding stoichiometry of the interaction process to be two base pairs per alkaloid molecule. The energetics of the strong interaction was studied by isothermal titration and differential scanning calorimetric techniques that suggested the binding to be exothermic and favoured by both negative enthalpy and positive entropy changes. All these results, together with the Stern–Volmer quenching experiment in fluorescence, revealed the molecular details of the intercalation of coralyne into poly(A) duplex leading to its potential use as an agent in gene regulation in eukaryotic cells.
Starting Page	341
Ending Page	348
Page Count	8
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	4
Issue Number	4
Journal	Molecular BioSystems
DOI	10.1039/b716356h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Stoichiometry RNA interference RNA Enthalpy Binding constant Volmer Fluorescence Eukaryote Entropy Alkaloid Bioenergetics Gene Job plot Scatchard Absorption spectroscopy Isothermal process Strong interaction Circular dichroism Outline of biophysics Exothermic process Hypochromic anemia Interferon Optical rotation Titration Scatchard equation
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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