title: The aconitase of yeast. V. The reconstitution of yeast aconitase.
abstract: The apoenzyme of yeast aconitase [EC 4.2.1.3] was prepared by treatment of yeast aconitase with sodium mersalyl, followed by passage by passage of the reaction mixture through a column of Dowex A-1 and gel filtration on Sephadex G-25. The apoenzyme had no aconitase activity, but the active enzyme could be reconstituted by treatment of the apoenzyme with ferrous ions and sodium sulfide in the presence of 2-mercapto-ethanol. The reconstituted active enzyme was isolated by DEAE-Sephadex A-50 column chromatography and Sephadex G-100 gel filtration from the reaction mixture. The reconstituted enzyme was identical with the original untreated enzyme in terms of specific activity, iron content and spectral characteristics, but not in terms of labile sulfur content. A significant difference in visible spectra between the holo- and apoenzymes appeared to be due to the difference in iron and labile sulfur contents between the two proteins.
mesh_terms: D000154:Aconitate Hydratase / Q000378:metabolism; D001051:Apoenzymes / Q000378:metabolism; D002175:Candida / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D006836:Hydro-Lyases / Q000378:metabolism; D007700:Kinetics; D008634:Mersalyl / Q000494:pharmacology; D013053:Spectrophotometry; D013056:Spectrophotometry, Ultraviolet
chemical_list: D001051:Apoenzymes; D008634:Mersalyl; D006836:Hydro-Lyases; D000154:Aconitate Hydratase