Quantitative analysis of the oxidative DNA lesion, 2,2-diamino-4-(2-deoxy-beta-D-erythro-pentofuranosyl)amino]-5(2H)-oxazolone (oxazolone), in vitro and in vivo by isotope dilution-capillary HPLC-ESI-MS/MS.
Quantitative analysis of the oxidative DNA lesion, 2,2-diamino-4-(2-deoxy-beta-D-erythro-pentofuranosyl)amino]-5(2H)-oxazolone (oxazolone), in vitro and in vivo by isotope dilution-capillary HPLC-ESI-MS/MS.
复制标题
氧化性DNA病变的定量分析，2,2-二氨基-4-（2-脱氧-beta-d-d-erthro-戊酰基）氨基）氨基] -5（2H）-Oxazolone（oxazolone），体外和体内通过同位素稀释-Capillary HPLC-ESI-MS/MS。
DOI：
10.1093/nar/gkl596
复制
发表时间：
2006
影响因子：
14.9
通讯作者：
Tretyakova N
中科院分区：
文献类型：
--
作者：
Matter B;Malejka-Giganti D;Csallany AS;Tretyakova N
脱氧核糖核酸的主要氧化产物2，2-diamino-4-[(2-deoxy-β-d-erythro-pentofuranosyl)amino]-5(2H)-oxazolone(恶唑酮)既可以直接氧化DG，也可以作为中间体8-氧代-7，8-二氢-2‘-脱氧鸟苷(8-oxo-DG)的二次氧化产物。定点突变研究表明，恶唑酮是一种强烈的错配病变，诱发的∼突变是8-oxo-DG的10倍。虽然8-oxo-DG容易进一步氧化，但恶唑酮似乎是鸟嘌呤氧化的稳定最终产物，如果在体内形成，可能成为氧化应激导致DNA损伤的生物标志物。本研究建立了毛细管液相色谱-电喷雾电离串联质谱仪(HPLC-ESI-MS/MS)对生物来源DNA中8-oxo-DG和恶唑酮的定量分析方法。用相应的15N标记内标进行同位素稀释，实现了对酶DNA水解物中8-oxo-DG和恶唑酮的灵敏和特异的检测。这两种碱基加合物在小牛胸腺DNA中以剂量依赖的方式在核黄素存在的光氧化作用下形成。随着辐照时间的延长，恶唑酮的生成量持续增加，而8-oxo-DG的生成量在20min时达到最大值，说明8-oxo-DG被转化为二次氧化产物。这两种损伤都是在仔细监测条件下分离的大鼠肝脏DNA中发现的，以最大限度地减少人为氧化。保持高动物脂肪饮食的糖尿病大鼠和对照组大鼠的肝脏DNA中每107个鸟嘌呤含有2-6个恶唑酮分子，而同一样品中的8-oxo-DG含量在每106个鸟嘌呤3-8个加合物之间。恶唑酮在大鼠肝DNA中损伤的形成、在氧化剂存在下的相对稳定性和强大的错配特性表明，恶唑酮可能在氧化应激介导的突变中发挥作用。
A major DNA oxidation product, 2,2-diamino-4-[(2-deoxy-β-d-erythro-pentofuranosyl)amino]-5(2H)-oxazolone (oxazolone), can be generated either directly by oxidation of dG or as a secondary oxidation product with an intermediate of 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxo-dG). Site-specific mutagenesis studies indicate that oxazolone is a strongly mispairing lesion, inducing ∼10-fold more mutations than 8-oxo-dG. While 8-oxo-dG undergoes facile further oxidation, oxazolone appears to be a stable final product of guanine oxidation, and, if formed in vivo, can potentially serve as a biomarker of DNA damage induced by oxidative stress. In this study, capillary liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) methods were developed to enable quantitative analysis of both 8-oxo-dG and oxazolone in DNA from biological sources. Sensitive and specific detection of 8-oxo-dG and oxazolone in enzymatic DNA hydrolysates was achieved by isotope dilution with the corresponding 15N-labeled internal standards. Both nucleobase adducts were formed in a dose-dependent manner in calf thymus DNA subjected to photooxidation in the presence of riboflavin. While the amounts of oxazolone continued to increase with the duration of irradiation, those of 8-oxo-dG reached a maximum at 20 min, suggesting that 8-oxo-dG is converted to secondary oxidation products. Both lesions were found in rat liver DNA isolated under carefully monitored conditions to minimize artifactual oxidation. Liver DNA of diabetic and control rats maintained on a diet high in animal fat contained 2–6 molecules of oxazolone per 107 guanines, while 8-oxo-dG amounts in the same samples were between 3 and 8 adducts per 106 guanines. The formation of oxazolone lesions in rat liver DNA, their relative stability in the presence of oxidants and their potent mispairing characteristics suggest that oxazolone may play a role in oxidative stress-mediated mutagenesis.
登录
查看更多内容
影响因子：
3.9
作者：
Greenberg, MM
通讯作者：
Greenberg, MM
影响因子：
14.9
作者：
Duarte, V;Gasparutto, D;Cadet, J
通讯作者：
Cadet, J
影响因子：
4.1
作者：
Gasparutto, D;Da Cruz, S;Cadet, J
通讯作者：
Cadet, J
影响因子：
3.3
作者：
Douki, T;Cadet, J
通讯作者：
Cadet, J
影响因子：
3.9
作者：
Hirakawa, K;Suzuki, H;Kawanishi, S
通讯作者：
Kawanishi, S