Receptor-mediated generation of an EDRF-like intermediate in a neuronal cell line detected by spin trapping techniques

Carmen M. Arroyo, Carlos Forray, Esam E. El-Fakahany, Gerald M. Rosen

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25 Scopus citations


We have studied receptor-mediated generation of an activator of soluble guanylate cyclase in cultured mouse neuroblastoma cells (clone N1E-115) by ESR/spin trapping spectroscopy. A spin adduct was detected during the activation of muscarinic receptors by carbamylcholine in the presence of the spin trap 3,5-dibromo 4-nitrosobenzene sulphonate (DBNBS). The spin adduct does not correspond to that originating from the free radical nitric oxide or hydroxylamine. The same adduct was generated in cytosol preparations from N1E-115 cells incubated with L-arginine, NADPH, in the presence of calcium. The use of isotopically labelled guanidino-N15-L-arginine supported the generation of a DBNBS spin trapped adduct originating from the guanidino moiety of L-arginine. Superoxide dismutase (SOD) stabilized the precursor of the spin adduct as well as the activator of soluble guanylate cyclase derived from L-arginine. Our results provide direct evidence for the receptor-mediated formation of a diffusible precursor of NO{up triangle, filled} derived from L-arginine.

Original languageEnglish (US)
Pages (from-to)1177-1183
Number of pages7
JournalBiochemical and Biophysical Research Communications
Issue number3
StatePublished - Aug 16 1990

Bibliographical note

Funding Information:
This researchw as supported by grants from NIH, (G.M.R.), NIH and U.S. Army ResearchO ffice (E.E.E.).


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