The effect on butyryl-CoA dehydrogenase of reagents specific for nucleophilic sulphur.
Removal of the green colour of butyryl-CoA dehydrogenase by reagents specific for nucleophilic sulphur is shown to involve chemical modification of the tightly bound CoA persulphide. 5,5'-Dithiobis-(2-nitrobenzoic acid) (Ellman's reagent) de-greens the enzyme essentially irreversibly, with a stoicheiometry of approx. 1 mol/mol of FAD. A compound separated by subsequent gel filtration is eluted at the same position as a CoA-thionitrobenzoate standard. The 35S-labelled distal sulphur atom of CoA persulphide is separated from this material. The enzyme remains fully active. Phenylmercuric acetate also de-greens the enzyme. The extent to which thiols restore the green colour declines with time. Gel filtration of mercurial-treated enzyme separates low-Mr material containing a CoA moiety, an extra S atom and a phenylmercury moiety. This material, added to yellow butyryl-CoA dehydrogenase with an excess of thiol, re-forms green enzyme, but it loses this ability on storage. The results are explicable if it is assumed that the thionitrobenzoate derivative of CoA persulphide loses the extra sulphur atom much more readily than does the phenylmercury derivative.
D044926:Butyryl-CoA Dehydrogenase; D002850:Chromatography, Gel; D003065:Coenzyme A / Q000378:metabolism; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology*; D044943:Fatty Acid Desaturases / Q000378:metabolism*; D009579:Nitrobenzoates / Q000494:pharmacology*; D010662:Phenylmercuric Acetate / Q000494:pharmacology*; D010663:Phenylmercury Compounds / Q000494:pharmacology*; D013057:Spectrum Analysis; D013455:Sulfur / Q000378:metabolism; D045854:Veillonellaceae / Q000201:enzymology
D009579:Nitrobenzoates; D010663:Phenylmercury Compounds; D013455:Sulfur; D004228:Dithionitrobenzoic Acid; D044943:Fatty Acid Desaturases; D044926:Butyryl-CoA Dehydrogenase; D010662:Phenylmercuric Acetate; D003065:Coenzyme A; D004229:Dithiothreitol