Intracellular glutathione mediates the denitrosylation of protein nitrosothiols in the rat spinal cord.
Protein S-nitrosothiols (PrSNOs) have been implicated in the pathophysiology of neuroinflammatory and neurodegenerative disorders. Although the metabolically instability of PrSNOs is well known, there is little understanding of the factors involved in the cleavage of S-NO linkage in intact cells. To address this issue, we conducted chase experiments in spinal cord slices incubated with S-nitrosoglutathione (GSNO). The results show that removal of GSNO leads to a rapid disappearance of PrSNOs (t(1/2) approximately 2 hr), which is greatly accelerated when glutathione (GSH) levels are raised with the permeable analogue GSH ethyl ester. Moreover, PrSNOs are stable in the presence of the GSH depletor diethyl maleate, indicating that GSH is critical for protein denitrosylation. Inhibition of GSH-dependent enzymes (glutathione S-transferase, glutathione peroxidase, and glutaredoxin) and enzymes that could mediate denitrosylation (alcohol dehydrogense-III, thioredoxin and protein disulfide isomerase) do not alter the rate of PrSNO decomposition. These findings and the lack of protein glutathionylation during the chase indicate that most proteins are denitrosylated via rapid transnitrosylation with GSH. The differences in the denitrosylation rate of individual proteins suggest the existence of additional structural factors in this process. This study is relevant to our recent discovery that PrSNOs accumulate in the central nervous system of patients with multiple sclerosis.
Romero
Jorge M
JM
Department of Cell Biology and Physiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico 87131, USA.
Bizzozero
Oscar A
OA
D000818:Animals; D015153:Blotting, Western; D004791:Enzyme Inhibitors / Q000494:pharmacology; D054477:Glutaredoxins / Q000037:antagonists & inhibitors; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors; D066298:In Vitro Techniques; D008297:Male; D008298:Maleates / Q000494:pharmacology; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D026422:S-Nitrosoglutathione / Q000378:metabolism; D026403:S-Nitrosothiols / Q000378:metabolism*; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism*; D013879:Thioredoxins / Q000037:antagonists & inhibitors
D004791:Enzyme Inhibitors; D054477:Glutaredoxins; D008298:Maleates; D011506:Proteins; D026403:S-Nitrosothiols; D013879:Thioredoxins; D026422:S-Nitrosoglutathione; D005979:Glutathione Peroxidase; D005982:Glutathione Transferase; D019704:Protein Disulfide-Isomerases; C014476:diethyl maleate; D005978:Glutathione