Detection of iron-sulfur center-containing subunits of mitochondrial NADH dehydrogenase by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by high-performance gel permeation chromatography.
Soluble NADH dehydrogenase resolved from Complex I of the mitochondrial electron-transfer chain was subjected to gel electrophoresis in the presence of sodium dodecyl sulfate at 4 degrees C, and then the gel was stained for iron with bathophenanthroline disulfonate and thioglycolic acid. The 23,000-dalton subunit was markedly stained, and the 51,000-dalton subunit was also stained, but only slightly. High-performance gel permeation chromatography using an eluant containing 0.1% sodium dodecyl sulfate also demonstrated that these subunits contain an iron-sulfur center: the elution pattern recorded by light absorption at 400 nm gave two peaks corresponding to the positions of the subunits.
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods; D003579:Cytochrome Reductases / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D007506:Iron-Sulfur Proteins / Q000302:isolation & purification*; D008667:Metalloproteins / Q000302:isolation & purification*; D008929:Mitochondria, Heart / Q000201:enzymology*; D009245:NADH Dehydrogenase / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate
D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D010446:Peptide Fragments; D012967:Sodium Dodecyl Sulfate; D003579:Cytochrome Reductases; D009245:NADH Dehydrogenase