A versatile new tool to quantify abasic sites in DNA and inhibit base excision repair.
A versatile new tool to quantify abasic sites in DNA and inhibit base excision repair.
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DOI：
10.1016/j.dnarep.2014.12.006
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发表时间：
2015-03
期刊：
影响因子：
3.8
通讯作者：
Bhagwat, Ashok S.
中科院分区：
文献类型：
--
作者：
Wei, Shanqiao;Shalhout, Sophia;Ahn, Young-Hoon;Bhagwat, Ashok S.
关键词：
许多内源性和外源性药物和细胞过程在DNA中产生基本位点（AP）。如果不进行修复，AP位点会导致突变、链断裂和细胞死亡。醛反应剂甲氧基胺与AP位点反应并阻断其修复。另一种烷氧胺，ARP，用生物素标记AP位点，并用于量化这些位点。我们将这两种能力结合成一种alkoxyamine， AA3，它与AP位点反应，具有比ARP更好的pH分布和反应活性。此外，AA3包含一个用于生物正交点击化学的炔功能，可用于将各种生化标签连接到AP位点。我们使用点击化学方法用生物素和荧光分子标记AP位点，而不使用蛋白质或酶。在生理pH下，AA3的反应性比ARP好，产率也比ARP高。它比ARP使用更简单，使用它可以降低背景值，提高AP站点检测的灵敏度。我们还发现AA3抑制碱基位点修复的第一个酶APE-1的程度与甲氧基胺大致相同。此外，AA3增强了烷基化剂甲基甲烷磺酸盐杀死人体细胞的能力，在这种联合化疗中比甲氧基胺更有效。
A number of endogenous and exogenous agents, and cellular processes create abasic (AP) sites in DNA. If unrepaired, AP sites cause mutations, strand breaks and cell death. Aldehyde-reactive agent methoxyamine reacts with AP sites and blocks their repair. Another alkoxyamine, ARP, tags AP sites with a biotin and is used to quantify these sites. We have combined both these abilities into one alkoxyamine, AA3, which reacts with AP sites with a better pH profile and reactivity than ARP. Additionally, AA3 contains an alkyne functionality for bioorthogonal click chemistry that can be used to link a wide variety of biochemical tags to AP sites. We used click chemistry to tag AP sites with biotin and a fluorescent molecule without the use of proteins or enzymes. AA3 has a better reactivity profile than ARP and gives much higher product yields at physiological pH than ARP. It is simpler to use than ARP and its use results in lower background and greater sensitivity for AP site detection. We also show that AA3 inhibits the first enzyme in the repair of abasic sites, APE-1, to about the same extent as methoxyamine. Furthermore, AA3 enhances the ability of an alkylating agent, methylmethane sulfonate, to kill human cells and is more effective in such combination chemotherapy than methoxyamine.
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影响因子：
7.2
作者：
Goodman MF;Woodgate R
通讯作者：
Woodgate R
影响因子：
1.3
作者：
Bennett, Samuel E.;Kitner, Joshua
通讯作者：
Kitner, Joshua
影响因子：
2.8
作者：
Cabelof, DC;Nakamura, J;Heydari, AR
通讯作者：
Heydari, AR
DOI：
10.3390/molecules16097980
发表时间：
2011-09-15
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Cavanagh BL;Walker T;Norazit A;Meedeniya AC
通讯作者：
Meedeniya AC
影响因子：
2.9
作者：
LINDAHL, T;NYBERG, B
通讯作者：
NYBERG, B