PMID: 20631Sep 1, 1977

Purification and properties of human erythrocyte pyrimidine 5'-nucleotidase

Proceedings of the National Academy of Sciences of the United States of America
J D TorranceE Beutler

Abstract

A 250,000-fold purification of pyrimidine 5'-nucleotidase from human erythrocytes has been achieved using a combination of DEAE-cellulose chromatography, ammonium sulfate fractionation, gel filtration, and isoelectric focusing. Polyacrylamide disc and starch gel electrophoresis of the purified material show two strong protein bands. On starch gel these bands exhibited pyrimidine 5'-nucleotidase activity. Two faint protein bands devoid of enzyme activity were also found in the case of polyacrylamide electrophoresis. The enzyme has a pH optimum at 7.5 and is most stable between pH 6 and 7.5. The enzyme has pI of 5.0 and a molecular weight of 28,000 by gel filtration. The Km of the purified enzyme was 10 muM, compared to 40 muM when measured in hemolysate. The higher Km in the hemolysate is due to the presence of an inhibitor. Inorganic phosphate was shown to be a competitive inhibitor of pyrimidine 5'-nucleotidase and inorganic phosphate in the hemolysate may be responsible for increasing the Km of the enzyme for the substrate cytidine monophosphate.

References

Oct 1, 1974·The Journal of Clinical Investigation·W N ValentineW S Adams
Jan 1, 1968·Advances in Clinical Chemistry·O Bodansky, M K Schwartz

Citations

Jul 1, 1978·Clinica Chimica Acta; International Journal of Clinical Chemistry·H A Buc, J C Kaplan
Jul 2, 1979·Clinica Chimica Acta; International Journal of Clinical Chemistry·H FujiiK Nomura
Apr 14, 2006·British Journal of Haematology·Alberto ZanellaGiovanna Valentini
Nov 1, 1982·Analytical Biochemistry·J D TorranceW Mills
Jun 1, 1982·Biochemical Medicine·E Beutler, C West
Aug 1, 1980·British Journal of Haematology·J D TorranceT Jenkins
Jan 1, 1985·Annals of the New York Academy of Sciences·D E Paglia, W N Valentine
Mar 9, 2010·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·Hongli ZhuThomas Ming Swi Chang
Oct 10, 2001·The Journal of Biological Chemistry·Zoran MinicGuy Hervé
Jan 1, 1980·Hemoglobin·W N Valentine, D E Paglia
Jul 1, 1985·American Journal of Hematology·S Miwa, H Fujii

Related Concepts

Erythrocytes
Eryhem
Hydrogen-Ion Concentration
Nucleotidases
Pyrimidine Nucleotides

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