Abstract
Purpose: To study deoxyribonucleic acid (DNA) shearing capability of copper(II) complex of dithiothreitol (DTT) and to fevaluate its potential application in cancer therapy. Methods: A parrot green complex was synthesized by grinding copper acetate monohydrate and DTT in 1:2 molar ratio in a mortar until no fumes of acetic acid were observed. The complex was characterized using attenuated total reflectance-Fourier transform infra-red (ATR-FTIR), and x-ray diffraction (XRD) techniques. Further information was also collected through Karl Fischer titration, thermogravimetric analysis (TGA) and (magnetic moment. Cleavage of DNA was determined by agarose gel electrophoresis. The gel was then stained, analyzed and photographed under ultraviolet (UV) light. Results: ATR-FTIR confirmed the formation of copper(II) complex with DTT by binding through thiol group based on the disappearance of the thiol (-SH) stretching peak at 2545 cm -1 . The crystalline structure was elucidated by a sharp intense peak at 38.520 in XRD spectrum while the octahedral geometry of complex was inferred from a magnetic moment of 1.72 B.M. The results for water content obtained by Karl Fischer titration and TGA revealed that water molecules are not part of the coordination sphere of the complex. Cleavage study of DNA showed that the complex completely sheared the circular DNA compared to pure DTT. Conclusion: Solvent free synthesis of Copper(II)-DTT complex has been successfully achieved, and an anhydrous complex with octahedral geometry obtained. The complex has a greater potential to shear DNA molecule than pure DTT. Keywords: DNA shearing, Copper(II) complex, Dithiothreitol, Attenuated total reflectance-Fourier transform infra-red, Karl Fischer titration, Magnetic moment
Highlights
D,L-Dithiothreitol, Cleland’s reagent, has vast applications including biochemical synthesis of thiol proteins, chemical peptide synthesis and the study of protein chemistry[1,2,3,4]
Deoxyribonucleic acid (DNA) shearing is an important phenomenon owing to its applications in the field of biology appear in the range of 3200 - 3400 cm-1
DNA shearing using copper(II) complex is superior to that using DTT
Summary
D,L-Dithiothreitol, Cleland’s reagent, has vast applications including biochemical synthesis of thiol proteins, chemical peptide synthesis and the study of protein chemistry[1,2,3,4]. DTT is found to be present in high concentration in samples of biomolecules which are studied as targets or models for biological heavy metal ion binding and reactivity. It is noted that DTT is a strong chelating agent [3], but only a few studies have been reported for its interactions with different metal ions [5,6,7,8,9,10]. Deoxyribonucleic acid (DNA) is observed to be a target for most of the anticancer and antiviral therapies [11,12]. Reedijk et al have investigated the complex [Cu(II) (pyrimol)Cl] having efficient self-activated DNA cleavage [14]
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