Abstract
Here we have examined the association of an aureolic acid antibiotic, chromomycin A3 (CHR), with Cu2+. CHR forms a high affinity 2:1 (CHR:Cu2+) complex with dissociation constant of 0.08 × 10−10 M2 at 25°C, pH 8.0. The affinity of CHR for Cu2+ is higher than those for Mg2+ and Zn2+ reported earlier from our laboratory. CHR binds preferentially to Cu2+ in presence of equimolar amount of Zn2+. Complex formation between CHR and Cu2+ is an entropy driven endothermic process. Difference between calorimetric and van’t Hoff enthalpies indicate the presence of multiple equilibria, supported from biphasic nature of the kinetics of association. Circular dichroism spectroscopy show that [(CHR)2:Cu2+] complex assumes a structure different from either of the Mg2+ and Zn2+ complex reported earlier. Both [(CHR)2:Mg2+] and [(CHR)2:Zn2+] complexes are known to bind DNA. In contrast, [(CHR)2:Cu2+] complex does not interact with double helical DNA, verified by means of Isothermal Titration Calorimetry of its association with calf thymus DNA and the double stranded decamer (5′-CCGGCGCCGG-3′). In order to interact with double helical DNA, the (antibiotic)2 : metal (Mg2+ and Zn2+) complexes require a isohelical conformation. Nuclear Magnetic Resonance spectroscopy shows that the Cu2+ complex adopts a distorted octahedral structure, which cannot assume the required conformation to bind to the DNA. This report demonstrates the negative effect of a bivalent metal upon the DNA binding property of CHR, which otherwise binds to DNA in presence of metals like Mg2+and Zn2+. The results also indicate that CHR has a potential for chelation therapy in Cu2+ accumulation diseases. However cytotoxicity of the antibiotic might restrict the use.
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Acknowledgments
We thank Prof. Siddhartha Roy, Director, Indian Institute of Chemical Biology (IICB), Kolkata for allowing us to access the central NMR facility and Mr. E. Padmanabhan for his technical assistance. We thank Prof. Toshiharu Hase, Director, Institute for Protein Research (IPR), Osaka University for inviting SL through the International Collaborative Research Programme with Professor Takao to perform the ESI–MS experiments. We also thank Ms. Parijat Majumder for critical reading of the manuscript. This work was supported by the intramural grant (Molecular Mechanism of Disease and Drug Action (MMDDA) project) [Grant number: 11-R&D-SIN-5.04] from Department of Atomic Energy, Govt. of India.
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10534_2011_9516_MOESM1_ESM.pdf
Kinetic parameters for the association of CHR with Cu2+. Normalised thermograms for the interaction of [(CHR)2:Cu2+] and [(CHR)2:Zn2+] complexes with the double stranded DNA decamer (5′-CCGGCGCCGG-3′) and thermodynamic parameters for the association of [(CHR)2:Zn2+] complex with the decamer. Supplementary material 1 (PDF 159 kb)
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Lahiri, S., Takao, T., Devi, P.G. et al. Association of aureolic acid antibiotic, chromomycin A3 with Cu2+ and its negative effect upon DNA binding property of the antibiotic. Biometals 25, 435–450 (2012). https://doi.org/10.1007/s10534-011-9516-4
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DOI: https://doi.org/10.1007/s10534-011-9516-4