PARTICULAR ABILITY OF LIVER P450S3A TO CATALYZE THE OXIDATION OF N(OMEGA)-HYDROXYARGININE TO CITRULLINE AND NITROGEN-OXIDES AND OCCURRENCE IN NO SYNTHASES OF A SEQUENCE VERY SIMILAR TO THE HEME-BINDING SEQUENCE IN P450S

Liver microsomes from rats pretreated with various inducers of P450 isoforms exhibit very different abilities to catalyze the oxidation of Nω- 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-aminoacid 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. © 1993 Academic Press, Inc.