Preparation and properties of S-nitroso-L-cysteine ethyl ester, an intracellular nitrosating agent.
In this report, a protocol for the preparation of the hydrochloride of S-nitroso-L-cysteine ethyl ester (SNCEE.HCl; 2) is presented. The synthesis of 2 has been targeted because S-nitroso-L-cysteine (SNC; 2b), which is extensively used for trans-S-nitrosation of thiol-containing proteins, has a limited ability of crossing cellular membranes. The nitrosothiol 2 was prepared via direct S-nitrosation of the hydrochloride of L-cysteine ethyl ester (CEE.HCl; 1a) with ethyl nitrite. 2 is relatively stable in crystal form and when neutralized to SNCEE (2a) in aqueous solutions treated with chelators of metal ions. Traces of metal ions, however, triggered the decomposition of 2a to nitric oxide and a S-centered radical, which were detected by ESR spectrometry. In contrast to 2b, 2a is a lipophilic compound that was taken up by human neutrophils. The latter process was paralleled by inhibition of the NADPH oxidase-dependent generation of superoxide anion radicals, presumably via reaction(s) of intracellular trans-S-nitrosation. Intracellular accumulation of S-nitrosothiols was observed with 2a but not with 2b. It is expected that the use of 2a will be advantageous when intracellular reactions of trans-S-nitrosation are to be studied.
Clancy
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Department of Biochemistry, Mount Sinai School of Medicine, New York, New York 10029, USA.
Cederbaum
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AI
Stoyanovsky
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DA
D003545:Cysteine / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D009603:Nitroso Compounds / Q000097:blood / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D013053:Spectrophotometry; D013481:Superoxides / Q000097:blood
D007202:Indicators and Reagents; D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; C429310:S-nitrosocysteine ethyl ester; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019255:NADPH Oxidases; D003545:Cysteine