Effect of superoxide dismutase on the stability of S-nitrosothiols.
S-Nitrosothiols formed from the nitric oxide (NO)-dependent S-nitrosation of thiol-containing proteins and peptides such as albumin and glutathione (GSH) have been implicated in the transport, storage, and metabolism of NO in vivo. Recent data suggest that certain transition metals enhance the decomposition of S-nitrosothiols in vitro. The objective of this study was to determine what effect Cu, Zn superoxide dismutase (CuZn-SOD) has on the stability of certain S-nitrosothiols such as S-nitrosoglutathione (GSNO) in vitro. We found that CuZn-SOD (20 microM) but not Mn-SOD in the presence of GSH catalyzed the decomposition of GSNO with a Vmax of 6.7 +/- 0.4 microM/min and a Km of 5.6 +/- 0.5 microM at 37 degreesC. Increasing GSH concentrations with respect to CuZn-SOD resulted in complete decomposition of GSNO at concentrations of GSH:SOD of 2:1. Increasing GSH concentrations further from 0.1 to 10 mM resulted in a concentration-dependent attenuation in GSNO decomposition suggesting that SOD-catalyzed decomposition of GSNO would be maximal at concentrations of GSH known to be present in extracellular fluids (e.g., plasma). The decomposition of GSNO by CuZn-SOD resulted in the sustained production of NO. We propose that GSH reduces enzyme-associated Cu2+ to Cu1+ which mediates the reductive decomposition of the S-nitrosothiol to yield free NO. We conclude that CuZn-SOD may represent an important physiological modulator of steady-state concentrations of low-molecular-weight S-nitrosothiols in vivo.
Jourd'heuil
D
D
Department of Molecular and Cellular Physiology, Louisiana State University Medical Center, Shreveport, Louisiana, 71130, USA.
Laroux
F S
FS
Miles
A M
AM
Wink
D A
DA
Grisham
M B
MB
D002384:Catalysis; D003545:Cysteine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009603:Nitroso Compounds / Q000737:chemistry* / Q000378:metabolism; D026422:S-Nitrosoglutathione; D026403:S-Nitrosothiols; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000378:metabolism
D009603:Nitroso Compounds; D026403:S-Nitrosothiols; C091665:S-nitrosoalbumin; D012710:Serum Albumin, Bovine; D026422:S-Nitrosoglutathione; C021315:S-nitrosocysteine; D013482:Superoxide Dismutase; D005978:Glutathione; D003545:Cysteine