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| MOLECULAR-CLONING AND FUNCTIONAL EXPRESSION OF AN INDUCIBLE NITRIC-OXIDE SYNTHASE FROM A MURINE MACROPHAGE CELL-LINE |
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| Author(s): LYONS CR, ORLOFF GJ, CUNNINGHAM JM |
| Source: JOURNAL OF BIOLOGICAL CHEMISTRY Volume: 267 Issue: 9 Pages: 6370-6374 Published: MAR 25 1992 |
| Times Cited: 834 References: 37 |
| Abstract: Macrophages activated by exposure to cytokines and/or to endotoxin produce nitric oxide (NO.), a free radical that is a mediator of the host response to infection. Activation induces the expression of nitric oxide synthase, the enzyme that catalyzes formation of NO. from L-arginine and molecular oxygen. We report the cloning of a cDNA encoding the inducible nitric oxide synthase from a murine macrophage cell line, RAW264.7, exposed to interferon-gamma and lipopolysaccharide. Oocytes injected with mRNA transcribed from this cDNA demonstrate arginine-dependent production of nitrite, a stable metabolite of NO.. Nitrite production is blocked by the enzyme inhibitor, N(G)-monomethylarginine, and is independent of calcium/calmodulin. RAW264.7 cells demonstrate rapid accumulation of the nitric oxide synthase-encoding mRNAs upon activation. Comparison of the deduced amino acid sequence to the calcium/calmodulin-dependent nitric oxide synthase previously purified (Bredt, D. S., and Snyder, S. H. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 682-685) and cloned (Bredt, D. S., Hwang, P. M., Glatt, C. E., Lowenstein, C., Reed, R. R., and Synder, S. H. (1991) Nature 351, 714-718) from rat brain identifies shared binding sites for the cofactors NADPH and flavins in the C-terminal half of both proteins and an additional conserved region near the N terminus that may recognize L-arginine and/or contribute to the active site. |
| Document Type: Article |
| Language: English |
Addresses:
1. BRIGHAM & WOMENS HOSP, RM 925, THORN BLDG, 75 FRANCIS ST, BOSTON, MA 02115 USA
2. BRIGHAM & WOMENS HOSP, DEPT MED, DIV HEMATOL, BOSTON, MA 02115 USA
3. HARVARD UNIV, SCH MED, BOSTON, MA 02115 USA |
| Publisher: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 |
| Subject Category: Biochemistry & Molecular Biology |
| IDS Number: HK318 |
| ISSN: 0021-9258 |
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