Electrochemiluminescent arrays for cytochrome P450-activated genotoxicity screening.: DNA damage from benzo[a]pyrene metabolites
Electrochemiluminescent arrays for cytochrome P450-activated genotoxicity screening.: DNA damage from benzo[a]pyrene metabolites
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DOI：
10.1021/ac061975q
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发表时间：
2007-03-01
影响因子：
7.4
通讯作者：
Rusling, James F.
中科院分区：
文献类型：
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作者：
Hvastkovs, Eli G.;So, Minjeong;Rusling, James F.
据报道，适用于遗传毒性筛选的阵列在薄膜斑点中产生细胞色素P450酶（CYP）的代谢产物。将含有DNA、各种人细胞色素P450和电化学发光（ECL）生成金属聚合物[Ru（bpy）（2）PVP10]（2+）的阵列点暴露于H2O2以激活酶。使用CCD照相机同时观察来自所有斑点的ECL。以苯并[a]芘为底物，对DNA损伤的酶活性顺序为：CYP1B1 > CYP1A2 > CYP1A1 > CYP2E1 >肌红蛋白，与它们的代谢活性顺序相同。因此，这些阵列估计不同酶产生遗传毒性代谢物的相对倾向。这是ECL阵列用于高通量体外遗传毒性筛选的首次演示。
Arrays suitable for genotoxicity screening are reported that generate metabolites from cytochrome P450 enzymes (CYPs) in thin-film spots. Array spots containing DNA, various human cyt P450s, and electrochemiluminescence (ECL) generating metallopolymer [Ru(bpy)(2)PVP10](2+) were exposed to H2O2 to activate the enzymes. ECL from all spots was visualized simultaneously using a CCD camera. Using benzo[a]pyrene as a test substrate, enzyme activity for producing DNA damage in the arrays was found in the order CYP1B1 > CYP1A2 > CYP1A1 > CYP2E1 > myoglobin, the same as the order of their metabolic activity. Thus, these arrays estimate the relative propensity of different enzymes to produce genotoxic metabolites. This is the first demonstration of ECL arrays for high-throughput in vitro genotoxicity screening.