LOCALIZATION AND PROPERTIES OF ALDEHYDE REDUCTASE

Publisher Summary This chapter discusses the localization and properties of aldehyde reductase. It presents the purification and the physico-chemical properties of human liver aldehyde reductase. Purification of human liver aldehyde reductase was achieved by passing crude liver homogenate over the following chromatography resins: DEAE-cellulose, Sephadex G-lOO, hydroxyl-apatite, and DEAE-Sephadex A-50, which all had been equilibrated against 10 mM Na-phosphate buffer pH 7.0. The Sephadex G-100 and hydroxylapatite columns were developed with the same buffer; from the ion exchange resins the enzyme was eluted by a gradient 10 to 100 mM Na-phosphate pH 7.0. Three independent methods were used to determine the molecular weight: SDS-gel electrophoresis, ultracentrifugation, and gel filtration on Sephadex G-l00. The values obtained by SDS-gel electrophoresis and gel filtration differ significantly. An unspecific adsorption to dextran could be the cause for the lower molecular weight found by gel filtration. Such a phenomenon was excluded by the use of Bio-Gel, which gave the same results as Sephadex.