Abstract
Publisher Summary Acylglycerone-phosphate reductase—a membrane-bound enzyme present in animal tissues—catalyzes the reduction of long-chain acyldihydroxyacetone phosphate (acyI-DHAP) or long-chain alkyl-DHAP by NADPH to the corresponding sn -glycerol 3-phosphate derivatives. In liver and other organs, the major fraction of the enzyme is present in peroxisomes. The enzyme has been solubilized and partially purified from Ehrlich ascites cell microsomes and purified to homogeneity from guinea pig liver peroxisomes. This chapter describes a method for the preparation of the homogeneous enzyme from guinea pig liver peroxisomes. The enzyme activity is assayed by measuring the radioactive lipid product formed after incubating alkyl-DHAP with B-[4- 3 H] NADPH. The purification of alkyldihydroxyacetone phosphate is discussed in the chapter. On sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, the purified enzyme shows a single protein band of relative mobility corresponding to 60,000 molecular weight. On gel filtration, in the presence of octylglucoside, the molecular weight of the enzyme is estimated at 75,000. The enzyme has a broad pH optimum between pH 6.5 and 7.5.
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