Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification
The formation of oxidative lesions arising from double stranded DNA damage is of major significance to chemical biology from the perspective of application to human health. The quantification of purine lesions arising from γ-radiation-induced hydroxyl radicals (HO•) has been the subject of numerous...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2015-03-01
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| Series: | Frontiers in Chemistry |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fchem.2015.00018/full |
| _version_ | 1819078072488427520 |
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| author | Michael A. Terzidis Carla eFerreri Chryssostomos eChatgilialoglu Chryssostomos eChatgilialoglu |
| author_facet | Michael A. Terzidis Carla eFerreri Chryssostomos eChatgilialoglu Chryssostomos eChatgilialoglu |
| author_sort | Michael A. Terzidis |
| collection | DOAJ |
| description | The formation of oxidative lesions arising from double stranded DNA damage is of major significance to chemical biology from the perspective of application to human health. The quantification of purine lesions arising from γ-radiation-induced hydroxyl radicals (HO•) has been the subject of numerous studies, with discrepancies on the measured 5',8-cyclo-2'-deoxyadenosine (cdA) and 5',8-cyclo-2'-deoxyguanosine (cdG) lesions reported by different groups. Recently we reported an ameliorative protocol for the analysis of DNA damage with quantitative determination of these lesions via isotope dilution liquid chromatography coupled with tandem mass spectrometry. Herein we applied this protocol for the quantification of these tandem-type purine lesions along with 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dG) and 7,8-dihydro-8-oxo-2'-deoxyadenosine (8-oxo-dA) in single and double stranded DNA, generated during DNA exposure to diffusible HO• radicals in the absence or presence of physiological levels of oxygen. The cdA and cdG lesions in absence of oxygen were found ~2 times higher in single than double stranded DNA, with 5'R being ~6.5 and ~1.5 times more predominant than 5'S in cdG and cdA, respectively. Interestingly, in the presence of 5% molecular oxygen the R/S ratios are retained with substantially decreased yields for cdA and cdG, whereas 8-oxo-dA and 8-oxo-dG remain nearly constant. The overall lesion formation follows the order: 8-oxo-dG >> 8-oxo-dA > 5'R-cdG > 5'R-cdA > 5'S-cdA > 5'S-cdG. By this method, there was a conclusive evaluation of radiation-induced DNA purine lesions. |
| first_indexed | 2024-12-21T19:07:17Z |
| format | Article |
| id | doaj.art-6f8078b9d8ea406d986f2510cc30eb87 |
| institution | Directory Open Access Journal |
| issn | 2296-2646 |
| language | English |
| last_indexed | 2024-12-21T19:07:17Z |
| publishDate | 2015-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Chemistry |
| spelling | doaj.art-6f8078b9d8ea406d986f2510cc30eb872022-12-21T18:53:17ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462015-03-01310.3389/fchem.2015.00018133046Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantificationMichael A. Terzidis0Carla eFerreri1Chryssostomos eChatgilialoglu2Chryssostomos eChatgilialoglu3Consiglio Nazionale delle RicercheConsiglio Nazionale delle RicercheConsiglio Nazionale delle RicercheN.C.S.R. “Demokritos”The formation of oxidative lesions arising from double stranded DNA damage is of major significance to chemical biology from the perspective of application to human health. The quantification of purine lesions arising from γ-radiation-induced hydroxyl radicals (HO•) has been the subject of numerous studies, with discrepancies on the measured 5',8-cyclo-2'-deoxyadenosine (cdA) and 5',8-cyclo-2'-deoxyguanosine (cdG) lesions reported by different groups. Recently we reported an ameliorative protocol for the analysis of DNA damage with quantitative determination of these lesions via isotope dilution liquid chromatography coupled with tandem mass spectrometry. Herein we applied this protocol for the quantification of these tandem-type purine lesions along with 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dG) and 7,8-dihydro-8-oxo-2'-deoxyadenosine (8-oxo-dA) in single and double stranded DNA, generated during DNA exposure to diffusible HO• radicals in the absence or presence of physiological levels of oxygen. The cdA and cdG lesions in absence of oxygen were found ~2 times higher in single than double stranded DNA, with 5'R being ~6.5 and ~1.5 times more predominant than 5'S in cdG and cdA, respectively. Interestingly, in the presence of 5% molecular oxygen the R/S ratios are retained with substantially decreased yields for cdA and cdG, whereas 8-oxo-dA and 8-oxo-dG remain nearly constant. The overall lesion formation follows the order: 8-oxo-dG >> 8-oxo-dA > 5'R-cdG > 5'R-cdA > 5'S-cdA > 5'S-cdG. By this method, there was a conclusive evaluation of radiation-induced DNA purine lesions.http://journal.frontiersin.org/Journal/10.3389/fchem.2015.00018/fullDNA DamageFree RadicalsLC-MS/MSgamma radiation8-oxo-dGcyclopurines |
| spellingShingle | Michael A. Terzidis Carla eFerreri Chryssostomos eChatgilialoglu Chryssostomos eChatgilialoglu Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification Frontiers in Chemistry DNA Damage Free Radicals LC-MS/MS gamma radiation 8-oxo-dG cyclopurines |
| title | Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification |
| title_full | Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification |
| title_fullStr | Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification |
| title_full_unstemmed | Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification |
| title_short | Radiation-induced formation of purine lesions in single and double stranded DNA: Revised quantification |
| title_sort | radiation induced formation of purine lesions in single and double stranded dna revised quantification |
| topic | DNA Damage Free Radicals LC-MS/MS gamma radiation 8-oxo-dG cyclopurines |
| url | http://journal.frontiersin.org/Journal/10.3389/fchem.2015.00018/full |
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