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Characterization of nitric oxide–stimulated ADP‐ribosylation of various proteins from the mouse macrophage cell line ANA‐1 using sodium nitroprusside and the novel nitric oxide‐donating compound diethylamine dinitric oxide
Author(s) -
Sheffler Lynn A.,
Wink David A.,
Melillo Giovanni,
Cox George W.
Publication year - 1995
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.57.1.152
Subject(s) - sodium nitroprusside , nitric oxide , adp ribosylation , biology , nitric oxide synthase , diethylamine , cholera toxin , autocrine signalling , biochemistry , microbiology and biotechnology , chemistry , nad+ kinase , enzyme , endocrinology , receptor , organic chemistry
We examined the ability of nitric oxide (NO) to stimulate the ADP‐ribosylation of proteins from the mouse macrophage cell line ANA‐1. To demonstrate a specific effect of NO, we used a novel compound named diethylamine dinitric oxide (DEA/NO; l,l‐diethyl‐2‐hydroxy‐2‐nitrosohydrazine, sodium salt; [Et 2 NN(O)NO]Na), which releases NO in aqueous solution at neutral pH. DEA/NO stimulated the ADP‐ribosylation of at least three cytosolic proteins (M r = 28,000, 33,000, and 39,000) from ANA‐1 macrophages. The effect of DEA/NO on the ADP‐ribosylation of the predominant target p39 was dose dependent (EC 50 = 80 μ M). Moreover, the effect of DEA/NO was attributed specifically to released NO rather than diethylamine or nitrite. Sodium nitroprusside (SNP) also stimulated the ADP‐ribosylation of cytosolic proteins from ANA‐1 mouse macrophages. However, SNP exhibited different time‐ and dose‐dependent effects on the modification of p39. NO synthesized via the activity of interferon‐γ plus lipopolysaccharide‐induced NO synthase also enhanced the ADP‐ribosylation of p39, confirming that the effects of DEA/NO and SNP could be attributed to NO or reactive nitrogen oxide species. Neither pertussis toxin nor cholera toxin stimulated the ADP‐ribosylation of p39; however, cholera toxin stimulated the ADP‐ribosylation of proteins with approximate molecular weights of 28,000 and 33,000. These data suggest that the induced expression of NO synthase in tumoricidal macrophages may be associated with autocrine and paracrine effects of NO that include the ADP‐ribosylation of various proteins. Moreover, these results indicate that DEA/NO and related compounds may be useful as pharmacologic tools for investigating the effects of NO and reactive nitrogen oxide species on macrophages. J. Leukoc. Biol. 57: 152–159; 1995.