Molecular and catalytic properties of a butyrylesterase from human red cells and brain.
A butyrylesterase from human red cells was prepared to homogeneity using DEAE-cellulose, Ultrogel ACA-34, DEAE-Sephacel, and precipitation with 1.5 M (NH4)2SO4. The yield was 25-35% relative to the enzyme activity of the hemolysate. Because of its preference for butyric acid esters the enzyme was designated a butyrylesterase. With alpha-naphthyl butyrate the Km was 7.6 microM and the kcat, 48 s-1. The molecular weight was 340,000 and the subunit weight 85,000, indicating a tetrameric structure. The isoelectric pH was 4.0. The enzyme preparation did not contain cystine. Sialic acid or other carbohydrate components could not be detected. The enzyme was irreversibly inhibited by organophosphate esters and the second-order rate constant was 192 M-1 s-1 for diethyl p-nitrophenyl phosphate. For the brain enzyme the constant was 206 M-1 s-1. The enzyme was irreversibly inhibited by sulfhydryl reagents, indicating that the enzyme is a sulfhydryl-dependent serine esterase. The enzyme was identical to the butyrylesterase from human brain, and the two enzymes were immunochemically identical. An amino acid ester has been shown to be split at a higher rate than butyric acid esters; however, the specificity constant (kcat/Km) was lower for the amino acid ester than for the butyric acid ester. The enzyme did not exhibit amidase activity.
Højring
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Department of Biochemistry C, Faculty of Medicine, University of Copenhagen, Denmark.
Svensmark
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D000596:Amino Acids / Q000032:analysis; D001921:Brain / Q000201:enzymology*; D002241:Carbohydrates / Q000032:analysis; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000097:blood / Q000302:isolation & purification*; D002384:Catalysis; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D000596:Amino Acids; D002241:Carbohydrates; D013439:Sulfhydryl Reagents; D002265:Carboxylic Ester Hydrolases; C021487:butyrylesterase