Structural gene products of the murine Ah complex. Differences in ontogenesis and glucosamine incorporation between liver microsomal cytochromes P1-450 and P-448 induced by polycyclic aromatic compounds.

M. Negishi, N. M. Jensen, G. S. Garcia, D. Nebert
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引用次数: 36

Abstract

Antibodies against mouse-liver microsomal cytochromes P1-450 and P-448, two polycyclic aromatic inducible cytochromes, were previously developed [Negishi, M. and Nebert, D.W. (1979) J. Biol. Chem. 254, 11015-11023]. Liver microsomes from 3-methylcholanthrene-treated and phenobarbital-treated and control adult mice and 2,3,7,8-tetrachlorodibenzo-p-dioxin-treated adult and fetal mice were examined. Immunoprecipitable radioactivity was measured, following labeling with pyridoxal phosphate/NaB[3H]4 or with 125I-labeled p-aminosulfobenzoic acid/NaNO2 in vitro or with [3H]leucine, [14C]glucosamine, or [32P]O4 in vivo. (a) Induction of cytochrome P1-450 occurs developmentally earlier in gestation than induction of cytochrome P-448 when the mother is treated with polycyclic aromatic compounds. (b) There appears to be a basal form of cytochrome P-448 but no cytochrome P1-450 in control liver microsomes; inducibility of cytochrome P-448 thus ranges between 5--12-fold, whereas that of P1-450 is infinite. (c) Phenobarbital pretreatment induces no detectable P1-450 or P-448. (d) P-448 appears to be either greater in concentration than P1-450 in the membrane or more exposed than P1-450 on the microsomal membrane surface. (e) By the radioimmunoassay methods used, 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced P1-450 and P-448 in Ah-nonresponsive mice are indistinguishable from those in Ah-responsive mice; this is true in both the fetus and the adult. (f) Compared with P-448 expression, the expression of P1-450 is more closely associated with 3-methylcholanthrene-induced aryl hydrocarbon hydroxylase activity, and these two structural gene products are apparently regulated independently. (g) P-448 but not P1-450 appears to be a glycoprotein. These data illustrate further differences between two forms of polycyclic aromatic-inducible P-450 in mouse liver. Neither P1-450 nor P-448 appears to be a phosphoprotein. Neither anti-(P1-450) nor anti-(P-448) precipitates any forms of liver microsomal P-450 from beta-naphthoflavone-treated adult rabbits and, conversely, anti-LM4 (the antibody to rabbit liver microsomal P-450 form 4) does not precipitate any forms of liver microsomal P-450 from 3-methylcholanthrene-treated C57BL/6N mice.
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小鼠Ah复合体的结构基因产物。多环芳香族化合物诱导的肝微粒体细胞色素P1-450和P-448在个体发生和葡萄糖胺掺入方面的差异
针对小鼠肝微粒体细胞色素P1-450和P-448(两种多环芳香族诱导细胞色素)的抗体先前已被开发[Negishi, M. and Nebert, D.W. (1979) J. Biol.]。化学学报,2004,23(2):444 - 444。对3-甲基胆蒽处理和苯巴比妥处理的成年小鼠和2,3,7,8-四氯二苯并对二恶英处理的成年小鼠和胎儿小鼠的肝微粒体进行了检测。在体外用吡哆醛磷酸/NaB[3H]4或125i标记的对氨基磺酸/NaNO2或体内用[3H]亮氨酸、[14C]氨基葡萄糖或[32P]O4标记后,测量免疫可沉淀放射性。(a)细胞色素P1-450的诱导在妊娠期比细胞色素P-448的诱导发生得早。(b)在对照肝微粒体中似乎有细胞色素P-448的基础形式,但没有细胞色素P1-450;因此,细胞色素P-448的诱导率在5- 12倍之间,而P1-450的诱导率是无限的。(c)苯巴比妥预处理未诱导检测到P1-450或P-448。(d) P-448在膜上的浓度似乎高于P1-450,或在微粒体膜表面的暴露程度高于P1-450。(e)采用放射免疫测定方法,ah无应答小鼠体内2,3,7,8-四氯二苯并-对二恶英诱导的P1-450和P-448与ah应答小鼠体内的P1-450和P-448无法区分;胎儿和成人都是如此。(f)与P-448的表达相比,P1-450的表达与3-甲基胆蒽诱导的芳烃羟化酶活性的关系更为密切,这两种结构基因产物明显是独立调控的。(g) P-448是糖蛋白,P1-450不是。这些数据进一步说明了小鼠肝脏中两种形式的多环芳烃诱导P-450之间的差异。P1-450和P-448似乎都不是磷蛋白。抗(P1-450)和抗(P-448)都不能从β -萘黄酮处理的成年家兔中沉淀任何形式的肝微粒体P-450,相反,抗lm4(兔肝微粒体P-450形式4的抗体)也不能从3-甲基胆碱处理的C57BL/6N小鼠中沉淀任何形式的肝微粒体P-450。
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