Macrophage synthesis of nitrite, nitrate, and N-nitrosamines: precursors and role of the respiratory burst

Journal Article · Tue Sep 01 04:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.18.6369· OSTI ID:7143394

Iyengar, R; Stuehr, D J; Marletta, M A

The macrophage cell line RAW 264.7 when activated with Escherichia coli lipopolysaccharide and interferon-..gamma.. synthesized nitrite (NO/sub 2//sup -/) and nitrate (NO/sub 3//sup -/). Medium change after the activation showed that L-arginine was the only amino acid essential for this synthesis. D-Arginine would not substitute for L-arginine. Other analogues that could replace L-arginine were L-homoarginine, L-arginine methyl ester, L-arginamide, and the peptide L-arginyl-L-aspartate. L-Argininic acid, L-agmatine, L-ornithine, urea, L-citrulline, and ammonia were among the nonprecursors, while L-canavanine inhibited this L-arginine-derived NO/sub 2//sup -//NO/sub 3//sup -/ synthesis. When morpholine was added to the culture medium of the activated RAW 264.7 macrophages, N-nitrosation took place, generating N-nitrosomorpholine. GC/MS experiments using L-(guanido-/sup 15/N/sub 2/)arginine established that the NO/sub 2//sup -//NO/sub 3//sup -/ and the nitrosyl group of N-nitrosomorpholine were derived exclusively from one or both of the terminal guanido nitrogens of arginine. Chromatographic analysis showed that the other product of the L-arginine synthesis of NO/sub 2//sup -//NO/sub 3//sup -/ was L-citrulline. The role of the respiratory burst in NO/sub 2//sup -//NO/sub 3//sup -/ synthesis was examined using the macrophage cell lines J774.16 and J774 C3C. Both cell lines synthesized similar amounts of NO/sub 2//sup -//NO/sub 3//sup -/. However, J774 C3C cells do not produce superoxide and hence do not exhibit the respiratory burst. Additional experiments also ruled out the involvement of the respiratory burst in NO/sub 2//sup -//NO/sub 3//sup -/ synthesis.

• Research Organization: Massachusetts Institute of Technology, Cambridge (USA)
• OSTI ID: 7143394
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Journal Issue: 18 Vol. 84:18; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ARGININE
BIOCHEMISTRY
BIOSYNTHESIS
CARBOXYLIC ACIDS
CHEMISTRY
CONNECTIVE TISSUE CELLS
HYDROGEN COMPOUNDS
HYDROGEN PEROXIDE
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
MACROPHAGES
MAMMALS
MICE
NITRATES
NITRITES
NITROGEN 15
NITROGEN COMPOUNDS
NITROGEN ISOTOPES
NITROSAMINES
NITROSO COMPOUNDS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXYGEN COMPOUNDS
PEROXIDES
PHAGOCYTES
RESPIRATION
RODENTS
SOMATIC CELLS
STABLE ISOTOPES
SYNTHESIS
VERTEBRATES