Enantioselective cleavage of supercoiled plasmid DNA catalyzed by chiral macrocyclic lanthanide(III) complexes |
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Authors: | Kr??el Artur Lisowski Jerzy |
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Institution: | a Department of Protein Engineering, Faculty of Biotechnology, University of Wroc?aw, Tamka 2, 50-137 Wroc?aw, Polandb Department of Chemistry, University of Wroc?aw, 14 F. Joliot-Curie, 50-383 Wroc?aw, Poland |
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Abstract: | The enantiomers of the Sm (III), Eu (III) and Yb (III) complexes LnL(NO3)2](NO3) of a chiral hexaazamacrocycle were tested as catalysts for the hydrolytic cleavage of supercoiled plasmid DNA. The catalytic activity was remarkably enantioselective; while the LnLSSSS(NO3)2](NO3) enantiomers promoted the cleavage of plasmid pBR322 from the supercoiled form (SC) to the nicked form (NC), the LnLRRRR(NO3)2](NO3) enantiomers were inactive. Kinetics of plasmid DNA hydrolysis was also investigated by agarose electrophoresis and it indicated typical single-exponential cleavage reaction. The hydrolytic mechanism of DNA cleavage was confirmed by the successful ligation of hydrolysis product by T4 ligase. The NMR study of the solutions of the complexes in various buffers indicated that the complexes exist as monomeric cationic complexes LnL(H2O)3]3 + in slightly acidic solutions and as dimeric cationic complexes Ln2L2(μ-OH)2(H2O)2]4 + in slightly basic 8 mM solutions, with the latter form being a possible catalyst for hydrolysis of phosphodiester bonds. |
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Keywords: | DNA cleavage Dinuclear lanthanide complex Enantioselectivity Chiral macrocycle |
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