Dynamic state of S-nitrosothiols in human plasma and whole blood.
In the vasculature, nitrosothiols derived from the nitric oxide (NO)-mediated S-nitrosation of thiols play an important role in the transport, storage, and metabolism of NO. The present study was designed to examine the reactions that promote the decomposition, formation, and distribution of extracellular nitrosothiols in the circulation. The disappearance of these species in plasma and whole blood was examined using a high-performance liquid chromatography method to separate low- and high-molecular weight nitrosothiols. We found that incubation of S-nitrosocysteine (CySNO) or S-nitrosoglutathione (GSNO) with human plasma resulted in a rapid decomposition of these nitrosothiols such that <10% of the initial concentration was recovered after 10-15 min. Neither metal chelators (DTPA, neocuproine), nor zinc chloride (glutathione peroxidase inhibitor), acivicin (gamma-glutamyl transpeptidase inhibitor), or allopurinol (xanthine oxidase inhibitor) inhibited the decomposition of GSNO. With both CySNO and GSNO virtually all NO was recovered as S-nitrosoalbumin (AlbSNO), suggesting the involvement of a direct transnitrosation reaction. Electrophilic attack of the albumin-associated thiols by reactive nitrogen oxides formed from the interaction of NO with O(2) was ruled out because one would have expected 50% yield of AlbSNO. Similar results were obtained in whole blood. The amount of S-nitrosohemoglobin recovered in the presence of 10 microM GSNO or CySNO was less than 100 nM taking into consideration the detection limit of the assay used. Our results suggest that serum albumin may act as a sink for low-molecular-weight nitrosothiols and as a modulator of NO(+) transfer between the vascular wall and intraerythrocytic hemoglobin.
Jourd'heuil
D
D
Vascular Biology Research Group, Albany Medical College, Albany, NY 12208, USA. david-jourd'heuil@ccgateway.amc.edu
Hallén
K
K
Feelisch
M
M
Grisham
M B
MB
D000493:Allopurinol / Q000494:pharmacology; D001711:Biotransformation; D002614:Chelating Agents / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000031:analogs & derivatives / Q000097:blood; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000031:analogs & derivatives / Q000097:blood; D006801:Humans; D007555:Isoxazoles / Q000494:pharmacology; D008623:Mercaptoethanol; D009603:Nitroso Compounds / Q000097:blood*; D010949:Plasma / Q000737:chemistry; D026422:S-Nitrosoglutathione; D026403:S-Nitrosothiols; D012709:Serum Albumin / Q000378:metabolism; D017967:Zinc Compounds / Q000494:pharmacology
D002614:Chelating Agents; D002712:Chlorides; D004791:Enzyme Inhibitors; D007555:Isoxazoles; D009603:Nitroso Compounds; D026403:S-Nitrosothiols; D012709:Serum Albumin; D017967:Zinc Compounds; D026422:S-Nitrosoglutathione; D008623:Mercaptoethanol; D000493:Allopurinol; C032635:S-nitrosomercaptoethanol; C016837:zinc chloride; C021315:S-nitrosocysteine; D005978:Glutathione; D003545:Cysteine; C006303:acivicin