A novel phosphodiesterase I from the soluble fraction of erythrocytes.
A previously unrecognized erythrocyte phosphodiesterase I with activity against thymidine-5'-monophospho-p-nitrophenyl ester is described. The enzyme is present in the soluble fraction of the erythrocyte, and was purified about 500-fold by chromatography using DEAE-cellulose, followed by gel chromatography with Sephadex G-200. Erythrocyte phosphodiesterase I has a molecular weight of about 70 000, when fully active as a monomer. Its pI is 5.4 and the pH optimum is 8.5. The Km value for thymidine-5'-monophospho-p-nitrophenyl ester is rather high, about 4 mmol/l. The enzyme has a barely detectable nucleotide pyrophosphatase activity. It is extremely sensitive to SH-inhibitors such as N-ethyl-maleimide, p-chloromercuribenzoate and disulphides (a reversible 50% inhibition was obtained by cystamine, 0.01 mmol/l). It is a metalloenzyme with loosely bound metal, and is stimulated by Mg2+. This activation by Mg2+ is counteracted by Zn2+. Gel chromatography revealed that the enzyme is a monomer in the presence of Mg2+. When inhibited by Zn2+, it forms polymers that can be reconverted to the monomer by thiols. All of the above properties of the erythrocyte enzyme support the conclusion that it is different from plasma membrane phosphodiesterase I (oligonucleate 5'-nucleotidohydrolase, EC 3.1.4.1).
D002413:Cations, Divalent / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004492:Edetic Acid / Q000494:pharmacology; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007526:Isoelectric Point; D007527:Isoenzymes / Q000097:blood / Q000302:isolation & purification*; D008970:Molecular Weight; D009711:Nucleotides / Q000494:pharmacology; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000097:blood / Q000302:isolation & purification*; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D002413:Cations, Divalent; D007527:Isoenzymes; D009711:Nucleotides; D013438:Sulfhydryl Compounds; D004492:Edetic Acid; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I