Inhibitory effects of nitric oxide and nitrosative stress on dopamine-beta-hydroxylase.
Dopamine-beta-hydroxylase (DbetaH) is a copper-containing enzyme that uses molecular oxygen and ascorbate to catalyze the addition of a hydroxyl group on the beta-carbon of dopamine to form norepinephrine. While norepinephrine causes vasoconstriction following reflex sympathetic stimulation, nitric oxide (NO) formation results in vasodilatation via a guanylyl cyclase-dependent mechanism. In this report, we investigated the relationship between NO and DbetaH enzymatic activity. In the initial in vitro experiments, the activity of purified DbetaH was inhibited by the NO donor, diethylamine/NO (DEA/NO), with an IC(50) of 1 mm. The inclusion of either azide or GSH partially restored DbetaH activity, suggesting the involvement of the reactive nitrogen oxide species, N(2)O(3). Treatment of human neuroblastoma cells (SK-N-MC) with diethylamine/NO decreased cellular DbetaH activity without affecting their growth rate and was augmented by the depletion of intracellular GSH. Co-culture of the SK-N-MC cells with interferon-gamma and lipopolysaccharide-activated macrophages, which release NO, also reduced the DbetaH activity in the neuroblastoma cells. Our results are consistent with the hypothesis that nitrosative stress, mediated by N(2)O(3), can result in the inhibition of norepinephrine biosynthesis and may contribute to the regulation of neurotransmission and vasodilatation.
Zhou
X
X
Laboratory of Human Carcinogenesis, Radiation Biology Branch, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Espey
M G
MG
Chen
J X
JX
Hofseth
L J
LJ
Miranda
K M
KM
Hussain
S P
SP
Wink
D A
DA
Harris
C C
CC
D000311:Adrenal Glands / Q000201:enzymology; D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D004299:Dopamine beta-Hydroxylase / Q000037:antagonists & inhibitors*; D005978:Glutathione / Q000494:pharmacology; D006801:Humans; D006834:Hydrazines / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects / Q000502:physiology; D051379:Mice; D017628:Microglia / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D009447:Neuroblastoma; D009569:Nitric Oxide / Q000494:pharmacology*; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D009589:Nitrogen Oxides; D019810:Sodium Azide / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D019323:omega-N-Methylarginine / Q000494:pharmacology
D006834:Hydrazines; D008070:Lipopolysaccharides; D020030:Nitric Oxide Donors; D009589:Nitrogen Oxides; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D007371:Interferon-gamma; C084012:1,1-diethyl-2-hydroxy-2-nitrosohydrazine; D019810:Sodium Azide; D004299:Dopamine beta-Hydroxylase; D005978:Glutathione