title: Studies on D-tetrose metabolism. Crystallization and properties of D-erythrulose reductase from chicken liver.
abstract: D-Erythrulose reductase from chicken liver has been purified to homogeneity as judged by acrylamide gel electrophoresis and ultracentrifugation. The overall purification of the enzyme was 164-fold from a crude extract. The enzyme was crystallized from ammonium sulfate solution at pH 7.0 to give hexagonal plates. The molecular weight determined by sedimentation equilibrium analysis was 94,600 and that by SDS-polyacrylamide gel electrophoresis was 22,400, which suggests a tetrameric structure for the native enzyme. The enzyme was found to contain up to 3 molecules of NADP+ per enzyme; this high amount of NADP+ resulted in a higher absorption at 260 nm than at 280 nm. The extinction coefficient of the enzyme at 290 nm was found to be 4.0. The contents of various amino acids were very similar to those of the beef liver enzyme formerly crystallized in our laboratory. The isoelectric point of the enzyme determined by Ampholine isoelectric focusing was pH 6.43. The enzyme was shown to catalyze the reduction of D-erythrulose to D-threitol with the concomitant oxidation of NAD(P)H to NAD(P)+, and was highly specific to D-erythrulose with an apparent Km of 0.38 mM. NADH was less effective than NADPH and the Km's for NADH and NADPH were 67 micrometers and 7.9 micrometers, respectively. D-Threitol was slightly oxidized by the enzyme with either NADP+ or NAD+ as a cofactor at pH's 7.5 and 9.0.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002645:Chickens; D003460:Crystallization; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D013780:Tetroses / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000596:Amino Acids; D013780:Tetroses; C100073:erythrulose; D013401:Sugar Alcohol Dehydrogenases; C020117:D-erythrulose reductase