Anti-Tumor Chloroquine-Gold Nanocomposites and their Binding Interaction with Bovine Serum Albumin: Biophysical and Bioc
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Anti-Tumor Chloroquine-Gold Nanocomposites and their Binding Interaction with Bovine Serum Albumin: Biophysical and Biochemical Aspects of Protein Binding Prachi Joshi,1,2 $ Soumyananda Chakraborti,3 $ Jaime E. Ramirez-Vick,4 Z. A. Ansari,2 Virendra Shanker1, Pinak Chakrabarti,3* Surinder P. Singh,1,4 * 1
National Physical Laboratory, Dr. K. S. Krishnan Marg, New Delhi-110012, India. Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi110025, India. 3 Department of Biochemistry, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata-700054, India. 4 Engineering Science and Materials Department, University of Puerto Rico, Mayaguez, PR 00680, USA. *Address for correspondence: [email protected]; [email protected] $ Both the authors have equal contribution. 2
ABSTRACT We have conjugated chloroquine onto nano-sized, thiol-stabilized gold nanoparticles by using 1ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) / Nhydroxysulfosuccinimide (NHS) chemistry. The formation of gold nanoparticles was confirmed using optical spectra for characteristic surface plasmon band; the average size of gold nanoparticles was found to be 5-7 nm from electron microscopy measurements. The anti-tumor activity of prepared nanocomposite, vis-à-vis chloroquine itself, had been demonstrated using MCF-7 breast cancer cell line. To determine the binding affinity of gold-chloroquine nanocomposites to transport proteins present in blood serum, we studied the binding interaction of gold-chloroquine to bovine serum albumin (BSA), the most abundant plasma protein. The binding was studied by using isothermal titration calorimetry and fluorescence spectroscopy and was analyzed in terms of binding constant, entropy and enthalpy change. The gold-chloroquine nanocomposites were found to interact efficiently with BSA and fluorescence quenching experiments involving Trp212 suggests that the nanocomposites bind at site I of BSA. INTRODUCTION Designing of hybrid nanocarrier systems capable of carrying DNA, proteins, drugs and other biomolecules for various biomedical applications has attracted a great attention in recent years.1 As drug delivery vehicles, gold nanoparticles have shown their potential with improve bioavailability, efficiency and specificity of pharmaceutical drugs, in particular anticancer agents.2 One of the reason for the choice of gold nanoparticles as drug delivery vehicles is their inert and nontoxic nature, ease in functionalization with desired ligands.3 Chloroquine is a wellknown inexpensive 9-aminoquinolone used as an anti-malarial drug. In addition to treating malaria, chloroquine is also effective as an antiviral agent against influenza and HIV-1.4 In recent years, metal (i.e., Au, Zn) complexed chloroquine has been used against drug-resistant malaria strains and as an anti-inflammatory agent.5 Chloroquine has also been shown to display anticancer activity and cause enhancement of the potency of other known drugs.6,7 In inorganic medicinal chemistry, cisplatin and other platinum a
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