Interaction of rhodanese with mitochondrial NADH dehydrogenase.
NADH dehydrogenase is an iron-sulfur flavoprotein which is isolated and purified from Complex I (mitochondrial NADH: ubiquinone oxidoreductase) by resolution with NaClO4. The activity of the enzyme (followed as NADH: 2-methylnaphthoquinone oxidoreductase) increases linearly with protein concentration (in the range between 0.2 and 1.0 mg/ml) and decreases with aging upon incubation on ice. In the present work a good correlation was found between enzymic activity and labile sulfide content, at least within the limits of sensitivity of the assays employed. Rhodanese (thiosulfate: cyanide sulfurtransferase (EC 2.8.1.1) purified from bovine liver mitochondria was shown to restore, in the presence of thiosulfate, the activity of the partly inactivated NADH dehydrogenase. Concomitantly, sulfur was transferred from thiosulfate to the flavoprotein and incorporated as acid-labile sulfide. Rhodanese-mediated sulfide transfer was directly demonstrated when the reactivation of NADH dehydrogenase was performed in the presence of radioactive thiosulfate (labeled in the outer sulfur) and the 35S-loaded flavoprotein was re-isolated by gel filtration chromatography. The results indicated that the [35S]sulfide was inserted in NADH dehydrogenase and appeared to constitute the structural basis for the increase in enzymic activity.
D000818:Animals; D002417:Cattle; D003579:Cytochrome Reductases / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology*; D016660:NAD(P)H Dehydrogenase (Quinone); D009245:NADH Dehydrogenase / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D011808:Quinone Reductases / Q000378:metabolism*; D013466:Sulfurtransferases / Q000378:metabolism*; D013884:Thiosulfate Sulfurtransferase / Q000378:metabolism*
D009247:NADH, NADPH Oxidoreductases; D003579:Cytochrome Reductases; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases; D009245:NADH Dehydrogenase; D013466:Sulfurtransferases; D013884:Thiosulfate Sulfurtransferase