Vol. 63, Issue 5, 1148-1158, May 2003

Role of S-Nitrosation of Cysteine Residues in Long-Lasting Inhibitory Effect of Nitric Oxide on Arterial Tone

Jacicarlos L. Alencar, Irina Lobysheva, Michel Geffard, Mamadou Sarr, Christa Schott, Valérie B. Schini-Kerth, Françoise Nepveu, Jean-Claude Stoclet, and Bernard Muller

Faculté de Pharmacie, Pharmacologie & Physico-Chimie, Université Louis Pasteur (Centre National de la Recherche Scientifique Unité Mixte Recherche 7034), 67401 Illkirch, France (J.L.A., M.S., C.S., V.B.S-K., J.-C. S, B.M.); Laboratório de Tecnologia Farmacêutica, Departamento de Ciências Básicas da Saúde, Universidade Federal da Paraíba, Campina Grande, Brazil (J.L.A.); Faculté des Sciences Pharmaceutiques, Laboratoire Pharmacophores Redox Phytochimie and Radiobiologie (EA-3030), Université Paul Sabatier, 31062 Toulouse, France (I.L., F.N.); Physique des Interactions Ondes-Matières (École Pratique des Hautes Études-Physique des Interactions Ondes Matiéres), 33600 Pessac, France (M.G.)

S-Nitrosation of cysteine residues plays an important role in nitric oxide (NO) signaling and transport. The aim of the present study was to investigate the role of S-nitrosothiols as a storage form of NO, which may account for the long-lasting effects in the vasculature. Rat aorta exposed to S-nitrosoglutathione (GSNO) displayed, even after washout of the drug, a persistent increase in cysteine-NO residues (detected by immunostaining using an antiserum that selectively recognized S-nitrosoproteins) and in NO content (detected by NO spin-trapping), a persistent attenuation of the effect of vasoconstrictors, and a relaxant response upon addition of low molecular weight (LMW) thiols. Rat mesenteric and porcine coronary artery exposed in vitro to GSNO, as well as aorta and mesenteric arteries removed from rats treated in vivo with GSNO, displayed similar modifications of contraction. In isolated aorta exposed to GSNO, the decrease of the contractile response and the relaxant effect of LMW thiols were both blunted by NO scavengers (oxyhemoglobin or 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) or by a cyclic GMP-dependent protein kinase inhibitor (Rp-8-bromoguanosine-3',5'-cyclic monophosphorothioate). In these arteries, mercuric chloride (which cleaves the cysteine-NO bond) exerted a transient relaxation, completely abolished the one of LMW thiols, and blunted the increase in cysteine-NO residues and NO content. Together, these data support the idea that S-nitrosation of cysteine residues is involved in long-lasting effects of NO on arterial tone. They suggest that S-nitrosation of tissue thiols is a mechanism of formation of local NO stores from which biologically active NO can subsequently be released.