Sheep brain glutathione reductase: purification and general properties

Sheep brain glutathione reductase was purified about 11,000-fold with an overall yield of 40%. The method included ammonium sulphate fractionation, heat denaturation, 2',5'-ADP Sepharose 4B and Sephadex G-200 chromatography steps. Specific activity at the final step was 193 IU/mg. The Mr of the enzyme was found to be 116,000 by gel filtration chromatography. On SDS-PAGE, two identical subunits of Mr 64,000 were obtained. From the spectral data, about 2 mol FAD per mol of enzyme were calculated.