Pterins inhibit nitric oxide synthase activity in rat alveolar macrophages.
1. The synthesis of nitrite and citrulline from L-arginine by immune-stimulated rat alveolar macrophages and the modulation of this synthesis were studied. 2,4-Diamino-6-hydroxypyrimidine (DAHP), 6R-5,6,7,8-tetrahydro-L-biopterin (BH4) and L-sepiapterin were potent inhibitors of the recombinant interferon-gamma induced production of nitrogen oxides in intact cultured cells with I50 values for BH4 and L-sepiapterin of approximately 10 microM. They were equally effective in inhibiting the induced production of citrulline. This inhibitory effect was concentration-dependent for all three modulators investigated. 2. The inhibitory effects were not dependent on incubation times of either 24 or 48 h, on the immune-stimulus used (lipopolysaccharide, interferon-gamma), or whether these stimuli were added during or after the induction period. 3. Pterin-6-carboxylic acid (PCA), which cannot be converted into BH4, and methotrexate (MTX), which inhibits dihydrofolatereductase but not de novo biosynthesis of BH4, did not change the production of nitrite. 4. The data indicate that DAHP, an inhibitor of the de novo biosynthesis of the co-factor BH4, blocks the nitric oxide synthase activity in intact cells. Since the pterins BH4 and L-sepiapterin blocked the L-arginine dependent production of nitrite and citrulline, the activity of nitric oxide synthase in phagocytic cells may be regulated by metabolic endproducts of the de novo biosynthesis of BH4.
D000594:Amino Acid Oxidoreductases / Q000037:antagonists & inhibitors*; D000818:Animals; D001120:Arginine / Q000378:metabolism; D001708:Biopterins / Q000031:analogs & derivatives* / Q000494:pharmacology; D002478:Cells, Cultured; D002956:Citrulline / Q000378:metabolism; D007042:Hypoxanthines / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000201:enzymology; D008297:Male; D019001:Nitric Oxide Synthase; D009573:Nitrites / Q000378:metabolism; D011621:Pteridines / Q000494:pharmacology*; D011622:Pterins; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins
D007042:Hypoxanthines; D009573:Nitrites; D011621:Pteridines; D011622:Pterins; D011994:Recombinant Proteins; D001708:Biopterins; D002956:Citrulline; D007371:Interferon-gamma; D001120:Arginine; C016727:sepiapterin; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; C017202:2,4-diaminohypoxanthine; C003402:sapropterin