Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Jan;81(2):588–592. doi: 10.1073/pnas.81.2.588

Identification of thymidine nucleotidase and deoxyribonucleotidase activities among normal isozymes of 5'-nucleotidase in human erythrocytes.

D E Paglia, W N Valentine, R A Brockway
PMCID: PMC344724  PMID: 6320196

Abstract

The persistence of normal thymidine nucleotidase (ThyNase) activity in subjects with pyrimidine nucleotidase (PyrNase) deficiency suggested the possible existence of separate isozymes in normal human erythrocytes. This hypothesis was confirmed by studies of PyrNase-deficient individuals from five unrelated families. Erythrocytes deficient in PyrNase retained normal activity of an enzyme system preferentially active at pH 6.2 with a variety of 2'-deoxyribonucleoside 5'-monophosphate substrates, including those of uridine, thymidine, and cytidine. Lesser activities were observed with the corresponding ribonucleotides. Normal control hemolysates were also found capable of effectively dephosphorylating purine nucleotides (dAMP greater than AMP) when pH was lowered sufficiently from the pH 7.4-8.0 region commonly used in conventional assays. Variations in substrate specificity, pH optima, kinetics, and sensitivity to inactivation by Pb2+ indicated the existence of multiple 5'-nucleotidase isozymes in normal erythrocytes: PyrNase and deoxyribonucleotidase(s) that might function physiologically in the conversion of DNA-derived nucleotides to diffusible nucleosides. Evolution of such a unique 5'-nucleotidase suggests that normal erythroblast maturation and nuclear extrusion is accompanied by a degree of karyolysis sufficient to require dephosphorylation and clearance of DNA degradation products.

Full text

PDF
588

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Awai M., Okada S., Takebayashi J., Kubo T., Inoue M., Seno S. Studies on the mechanism of denucleation of the erythroblast. Acta Haematol. 1968 Apr;39(4):193–202. doi: 10.1159/000208962. [DOI] [PubMed] [Google Scholar]
  2. Beutler E., Blume K. G., Kaplan J. C., Löhr G. W., Ramot B., Valentine W. N. International Committee for Standardization in Haematology: recommended methods for red-cell enzyme analysis. Br J Haematol. 1977 Feb;35(2):331–340. doi: 10.1111/j.1365-2141.1977.tb00589.x. [DOI] [PubMed] [Google Scholar]
  3. Beutler E. Red cell enzyme defects as nondiseases and as diseases. Blood. 1979 Jul;54(1):1–7. [PubMed] [Google Scholar]
  4. Fujii H., Nakashima K., Miwa S., Nomura K. Electrophoretic and kinetic studies of a mutant red cell pyrimidine 5'-nucleotidase. Clin Chim Acta. 1979 Jul 2;95(1):89–92. doi: 10.1016/0009-8981(79)90340-1. [DOI] [PubMed] [Google Scholar]
  5. Ishida Y., Fujii H., Miwa S. Pyrimidine 5'-nucleotidase deficiency: studies of five cases in two Japanese families. Hemoglobin. 1980;4(5-6):683–691. doi: 10.3109/03630268008997737. [DOI] [PubMed] [Google Scholar]
  6. Ishida Y., Miwa S., Miura Y., Kibe A. Electrophoretic and kinetic studies of human erythrocytes deficient in pyrimidine 5'-nucleotidase. Clin Chim Acta. 1980 Dec 8;108(2):285–291. doi: 10.1016/0009-8981(80)90015-7. [DOI] [PubMed] [Google Scholar]
  7. PEASE D. C. An electron microscopic study of red bone marrow. Blood. 1956 Jun;11(6):501–526. [PubMed] [Google Scholar]
  8. Paglia D. E., Valentine W. N. Characteristics of a pyrimidine-specific 5'-nucleotidase in human erythrocytes. J Biol Chem. 1975 Oct 25;250(20):7973–7979. [PubMed] [Google Scholar]
  9. Paglia D. E., Valentine W. N., Dahlgren J. G. Effects of low-level lead exposure on pyrimidine 5'-nucleotidase and other erythrocyte enzymes. Possible role of pyrimidine 5'-nucleotidase in the pathogenesis of lead-induced anemia. J Clin Invest. 1975 Nov;56(5):1164–1169. doi: 10.1172/JCI108192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Paglia D. E., Valentine W. N., Fink K. Lead poisoning. Further observations on erythrocyte pyrimidine-nucleotidase deficiency and intracellular accumulation of pyrimidine nucleotides. J Clin Invest. 1977 Dec;60(6):1362–1366. doi: 10.1172/JCI108896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Paglia D. E., Valentine W. N. Hereditary and acquired defects in the pyrimidine nucleotidase of human erythrocytes. Curr Top Hematol. 1980;3:75–109. [PubMed] [Google Scholar]
  12. Paglia D. E., Valentine W. N., Keitt A. S., Brockway R. A., Nakatani M. Pyrimidine nucleotidase deficiency with active dephosphorylation of dTMP: evidence for existence of thymidine nucleotidase in human erythrocytes. Blood. 1983 Nov;62(5):1147–1149. [PubMed] [Google Scholar]
  13. Rosa R., Rochant H., Dreyfus B., Valentin C., Rosa J. Electrophoretic and kinetic studies of human erythrocytes deficient in pyrimidine 5'-nucleotidase. Hum Genet. 1977 Sep 22;38(2):209–215. doi: 10.1007/BF00527404. [DOI] [PubMed] [Google Scholar]
  14. Shinohara K., Tanaka K. R. Kinetic and electrophoretic studies of human erythrocytes deficient in pyrimidine 5'-nucleotidase. Hum Genet. 1979 Sep 2;51(1):107–111. doi: 10.1007/BF00278299. [DOI] [PubMed] [Google Scholar]
  15. Simpson C. F., Kling J. M. The mechanism of denucleation in circulating erythroblasts. J Cell Biol. 1967 Oct;35(1):237–245. doi: 10.1083/jcb.35.1.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Skutelsky E., Danon D. An electron microscopic study of nuclear elimination from the late erythroblast. J Cell Biol. 1967 Jun;33(3):625–635. doi: 10.1083/jcb.33.3.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Swallow D. M., Aziz I., Hopkinson D. A., Miwa S. Analysis of human erythrocyte 5'-nucleotidases in healthy individuals and a patient deficient in pyrimidine 5'-nucleotidase. Ann Hum Genet. 1983 Jan;47(Pt 1):19–23. doi: 10.1111/j.1469-1809.1983.tb00966.x. [DOI] [PubMed] [Google Scholar]
  18. Torrance J. D., Whittaker D. A. Some properties of human erythrocyte pyrimidine 5'-nucleotidase. Enzyme. 1981;26(2):64–73. doi: 10.1159/000459152. [DOI] [PubMed] [Google Scholar]
  19. Valentine W. N., Fink K., Paglia D. E., Harris S. R., Adams W. S. Hereditary hemolytic anemia with human erythrocyte pyrimidine 5'-nucleotidase deficiency. J Clin Invest. 1974 Oct;54(4):866–879. doi: 10.1172/JCI107826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Valentine W. N., Paglia D. E., Fink K., Madokoro G. Lead poisoning: association with hemolytic anemia, basophilic stippling, erythrocyte pyrimidine 5'-nucleotidase deficiency, and intraerythrocytic accumulation of pyrimidines. J Clin Invest. 1976 Oct;58(4):926–932. doi: 10.1172/JCI108545. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES