The reaction of HOo radical with DNA is intensively studied both mechanistically and analytically for lesions formation. Several aspects related to the reaction paths of purine moieties with the formation of 5?,8-cyclopurines (cPu), 8-oxopurines (8-oxo-Pu) and their relationship are not well understood. In this study, we investigated the reaction of HOo radical with a 21-mer double-stranded oligodeoxynucleotide (ds-ODNs) in ?-irradiated aqueous solutions under various oxygen concentrations and accurately quantified the six purine lesions (i.e., four cPu and two 8-oxo-Pu) by LC-MS/MS analysis using isotopomeric internal standards. In the absence of oxygen 8-oxo-Pu lesions are only ~4 times more than cPu lesions. By increasing oxygen concentration, the 8-oxo-Pu and the cPu gradually increase and decrease, respectively, reaching a gap of ~130 times at 2.01×10-4 M of O2. Kinetic treatment of the data allows to estimate the C5? radical competition between cyclization and oxygen trapping in ds-ODNs, and lastly the rate constants of the four cyclization steps. Tailored computational studies by means of dispersion-corrected DFT calculations were performed on the CGC and TAT in their double-strand models for each cPu diastereoisomer along with the complete reaction pathways of the cyclization steps. Our findings reveal unheralded reaction mechanisms that resolve the long-standing issues with C5? radical cyclization in purine moieties of DNA sequences.
Oxygen Dependent Purine Lesions in Double-Stranded Oligodeoxynucleotides: Kinetic and Computational Studies Highlight the Mechanism for 5?,8-Cyplopurine Formation
Chryssostomos Chatgilialoglu;Annalisa Masi;
2020
Abstract
The reaction of HOo radical with DNA is intensively studied both mechanistically and analytically for lesions formation. Several aspects related to the reaction paths of purine moieties with the formation of 5?,8-cyclopurines (cPu), 8-oxopurines (8-oxo-Pu) and their relationship are not well understood. In this study, we investigated the reaction of HOo radical with a 21-mer double-stranded oligodeoxynucleotide (ds-ODNs) in ?-irradiated aqueous solutions under various oxygen concentrations and accurately quantified the six purine lesions (i.e., four cPu and two 8-oxo-Pu) by LC-MS/MS analysis using isotopomeric internal standards. In the absence of oxygen 8-oxo-Pu lesions are only ~4 times more than cPu lesions. By increasing oxygen concentration, the 8-oxo-Pu and the cPu gradually increase and decrease, respectively, reaching a gap of ~130 times at 2.01×10-4 M of O2. Kinetic treatment of the data allows to estimate the C5? radical competition between cyclization and oxygen trapping in ds-ODNs, and lastly the rate constants of the four cyclization steps. Tailored computational studies by means of dispersion-corrected DFT calculations were performed on the CGC and TAT in their double-strand models for each cPu diastereoisomer along with the complete reaction pathways of the cyclization steps. Our findings reveal unheralded reaction mechanisms that resolve the long-standing issues with C5? radical cyclization in purine moieties of DNA sequences.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
