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PMID:9226880
| Citation |
Kaspers, B, Gütlich, M, Witter, K, Lösch, U, Goldberg, M and Ziegler, I (1997) Coordinate induction of tetrahydrobiopterin synthesis and nitric oxide synthase activity in chicken macrophages: upregulation of GTP-cyclohydrolase I activity. Comp. Biochem. Physiol. B, Biochem. Mol. Biol. 117:209-15 |
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| Abstract |
Biosynthesis of nitric oxide (NO) and tetrahydrobiopterin (BH4) was investigated during cytokine-mediated activation of chicken macrophages. Monocyte derived macrophages and HD11 cells, a chicken macrophage cell line, constitutively synthesize BH4. Treatment of these cells with chicken macrophage activation factor (ChMAF) causes up to 10-fold increases of intracellular BH4 and of nitrite concentrations in the cell culture supernatant. Elevated BH4 levels correlate with an increase in GTP-cyclohydrolase I (GTP-CH) activity. Kinetic studies show a joint upregulation of GTP-CH activity and NO synthase activity first detectable 4 hr after stimulation. A corresponding increase in the mRNA for GTP-CH was detected by Northern blot analysis with a chicken GTP-CH specific cDNA probe. These results demonstrate that cytokine-induced BH4 synthesis by chicken macrophages is at least partially regulated through increased GTP-CH gene expression. The functional relevance of BH4 formation for NO production is shown by experiments using 2,4-diamino-6-hydroxypyrimidine (DAHP) as a specific inhibitor of GTP-CH. Monocyte derived macrophages stimulated in the presence of DAHP show a significant decrease in NO synthesis. The effect of DAHP was reversed by adding sepiapterin, which allows synthesis of BH4 through a salvage pathway. |
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| Keywords |
Animals; Biopterin/analogs & derivatives; Biopterin/biosynthesis; Blotting, Northern; Chickens; GTP Cyclohydrolase/genetics; GTP Cyclohydrolase/metabolism; Kinetics; Macrophages/enzymology; Nitric Oxide/biosynthesis; Nitric Oxide Synthase/metabolism; RNA, Messenger/metabolism |
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