The temporal relationships between P450E protein content, catalytic activity, and mRNA levels in the teleost Fundulus heteroclitus following treatment with beta-naphthoflavone.
The temporal relationships between P450E protein content, catalytic activity, and mRNA levels in the teleost Fundulus heteroclitus following treatment with beta-naphthoflavone.
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β-萘黄酮处理后硬骨鱼眼底异斜体中 P450E 蛋白含量、催化活性和 mRNA 水平之间的时间关系。
DOI：
10.1016/0003-9861(89)90319-6
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发表时间：
1989
影响因子：
3.9
通讯作者：
Stegeman,JJ
中科院分区：
文献类型：
--
作者：
Kloepper-Sams,PJ;Stegeman,JJ
一些海洋生物在接触化学物质后会发生P450诱导。通过检测单剂量β-萘黄酮组分对异阴蒂硬骨鱼肝脏同工酶P450E含量、酶活性及其基因表达水平的影响，探讨了3-甲基胆蒽(MC)的诱导方式。P450E是MC类化合物诱导的主要硬骨P450，是芳烃羟基酶(AHH)和乙氧基间苯二酚O-脱乙基酶(EROD)活性的催化剂。在为期20天的实验中，从第4天到第20天，接受BNF处理的动物的EROD活性都有所上升。免疫可检测到的P450E活性的增加与EROD活性的增加趋势相同，控制值始终较低，BNF处理的鱼至少增加了19倍。用抗P450E抗体沉淀肝脏蛋白体外翻译产物时，对照鱼没有检测到信号，而BNF处理的动物在6000mR条带中掺入了[~3H]亮氨酸。在较短期的实验中，BNF处理后EROD活性和P450E水平再次协调增加，免疫沉淀显示在处理后6h或更长时间的所有时间点，这些鱼的翻译产物P450E mRNA水平都有明显增加的趋势。经BNF处理的Fundulus的RNA与鲑鱼P450IA1基因的杂交也显示出随着时间的推移单条带的增加。P450E基因的表达先于P450E蛋白和酶活性升高约25h，但P450E基因表达迅速下降，在5d时达到对照水平，而蛋白质水平至少在13d内保持较高水平。结果支持一种假设，即转录增强参与了MC诱导的P450在鱼类中的诱导，但表明P450E的诱导也受到其他形式的调控。
P450 induction occurs in some marine organisms following chemical exposure. The mode of 3-methylcholanthrene (MC)-type induction was evaluated by examining hepatic isozyme P450E content, catalytic activity, and mRNA levels in the marine teleostFundulus heteroclitusafter exposure to a single dose of β-naphthoflavone (BNF). P450E is the major teleost P450 induced by MC-type compounds and is the catalyst for aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufinO-deethylase (EROD) activities. In a 20-day experiment, EROD activity was elevated in BNF-treated animals from Day 4 through Day 20. Increases in immunodetectable P450E showed the same trend as EROD activity, with consistently low control values and at least a 19-fold increase in the BNF-treated fish. Precipitation of liver RNAin vitrotranslation products with anti-P450E antibody gave no detectable signal from control fish, while the BNF-treated animals showed incorporation of [3H]leucine in a single 6,000Mrband. In a shorter term experiment, EROD activity and P450E levels were again coordinately increased in response to BNF treatment, and immunoprecipitation of translation products from these fish showed a clear trend of increased P450E mRNA levels for all time points 6 h or more post-treatment. Hybridization of RNA from BNF-treatedFunduluswith a trout P450IA1 cDNA also showed increases in a single band with time. The increases in P450E mRNA preceded increases in P450E protein and enzyme activity by about 25 h. However, P450E mRNA declined rapidly, reaching control levels by 5 days, while protein levels remained elevated for at least 13 days. The results support a hypothesis that transcriptional enhancement is involved in induction of MC-inducible P450s in fish, but indicate that P450E induction is also under other forms of regulatory control.
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