Chronic exposure to nitric oxide alters the free iron pool in endothelial cells: role of mitochondrial respiratory complexes and heat shock proteins.
The mechanisms of nitric oxide (NO) signaling include binding to the iron centers in soluble guanylate cyclase and cytochrome c oxidase and posttranslational modification of proteins by S-nitrosation. Low levels of NO control mitochondrial number in cells, but little is known of the impact of chronic exposure to high levels of NO on mitochondrial function in endothelial cells. The focus of this study is the interaction of NO with mitochondrial respiratory complexes in cell culture and the effect this has on iron homeostasis. We demonstrate that chronic exposure of endothelial cells to NO decreased activity and protein levels of complexes I, II, and IV, whereas citrate synthase and ATP synthase were unaffected. Inhibition of these respiratory complexes was accompanied by an increase in cellular S-nitrosothiol levels, modification of cysteines residues, and an increase in the labile iron pool. The NO-dependent increase in the free iron pool and inhibition of complex II was prevented by inhibition of mitochondrial protein synthesis, consistent with a major contribution of the organelle to iron homeostasis. In addition, inhibition of mitochondrial protein synthesis was associated with an increase in heat shock protein 60 levels, which may be an additional mechanism leading to preservation of complex II activity.
Ramachandran
Anup
A
Department of Pathology and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Ceaser
Erin
E
Darley-Usmar
Victor M
VM
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002701:Chloramphenicol / Q000494:pharmacology; D003545:Cysteine / Q000737:chemistry; D045222:Electron Transport Chain Complex Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D005293:Ferritins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009603:Nitroso Compounds / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry / Q000378:metabolism
D045222:Electron Transport Chain Complex Proteins; D006360:Heat-Shock Proteins; D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; D011500:Protein Synthesis Inhibitors; D011506:Proteins; C094210:2,2'-(hydroxynitrosohydrazono)bis-ethanamine; D009569:Nitric Oxide; D002701:Chloramphenicol; D005293:Ferritins; D007501:Iron; D003545:Cysteine