Renaud J P, Boucher J L, Vadon S, Delaforge M, Mansuy D
Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, URA 400, Université René Descartes, Paris, France.
Biochem Biophys Res Commun. 1993 Apr 15;192(1):53-60. doi: 10.1006/bbrc.1993.1380.
Liver microsomes from rats pretreated with various inducers of P450 isoforms exhibit very different abilities to catalyze the oxidation of N omega-hydroxy-L-arginine (NOHA) by NADPH and O2 with formation of citrulline and nitrogen oxides. Treatment of rats with dexamethasone, a classical inducer of P450 3A, leads to a spectacular 7-fold increase of the activity found for untreated rats, while induction by phenobarbital causes a much lower increase of this activity and induction by 3-methylcholanthrene or clofibrate decreases it. Specific inhibitors of P450s3A as troleandomycin and dihydroergotamine strongly inhibit NOHA oxidation whereas metyrapone, an inhibitor of other P450 subfamilies, was without effect. These data show the particular ability of P450s of the 3A subfamily to catalyze the second step of the oxidation of L-arginine by NO synthases (NOS). This analogy between NOSs and P450s3A is further substantiated by a protein sequence comparison which shows that a 9-amino acid segment present in all NOSs exhibits a strong similarity with the sequence mainly responsible for heme binding in P450s3A which is well conserved in all P450s. This segment contains all the structural factors which are thought to be crucial for heme binding in P450s.
用各种细胞色素P450同工型诱导剂预处理的大鼠肝脏微粒体,在催化Nω-羟基-L-精氨酸(NOHA)由NADPH和O2氧化生成瓜氨酸和氮氧化物方面表现出非常不同的能力。用地塞米松(一种经典的细胞色素P450 3A诱导剂)处理大鼠,会使未处理大鼠的活性显著增加7倍,而苯巴比妥诱导则使该活性增加幅度小得多,3-甲基胆蒽或氯贝丁酯诱导则使其降低。细胞色素P450s3A的特异性抑制剂如三乙酰竹桃霉素和双氢麦角胺强烈抑制NOHA氧化,而其他细胞色素P450亚家族的抑制剂美替拉酮则无作用。这些数据表明3A亚家族的细胞色素P450具有催化一氧化氮合酶(NOS)氧化L-精氨酸第二步反应的特殊能力。通过蛋白质序列比较进一步证实了NOS与细胞色素P450s3A之间的这种相似性,该比较表明所有NOS中存在的一个9个氨基酸的片段与细胞色素P450s3A中主要负责血红素结合的序列具有很强的相似性,该序列在所有细胞色素P450中都高度保守。该片段包含了被认为对细胞色素P450中血红素结合至关重要的所有结构因素。