Substrate specificity and pH sensitivity of deoxyribonucleotidase and pyrimidine nucleotidase activities in human hemolysates.
Residual 5'-nucleotidase activities in hemolysates from nine subjects with severe hereditary deficiency of pyrimidine nucleotidase (PyrNase) were compared to those in normal and reticulocyte-rich controls. Dephosphorylation rates of 12 potential ribo- and deoxyribomononucleotide substrates were measured as a function of pH. Data confirmed the existence of at least two isozymes of 5'-nucleotidase, PyrNase, and 2'-deoxy-5'-ribonucleotide phosphohydrolase (dNase) distinguishable by differences in maximal velocities, substrate preferences and restrictions, and pH optima. PyrNase was confirmed to be active principally with pyrimidine substrates (UMP = dCMP greater than CMP much greater than dTMP greater than dUMP) at a pH optimum of 7.5 +/- 0.1. dNase activity occurred with both purine and pyrimidine substrates and was maximal with deoxy analogs (dIMP much greater than dUMP greater than dGMP greater than dTMP = dAMP much greater than dCMP) at a pH optimum of 6.2, but slight cross-reactivity occurred with some nondeoxy substrates (IMP greater than GMP greater than UMP = XMP greater than CMP). PyrNase and dNase may be complementary systems that serve physiologically to clear the cytosol of RNA and DNA degradation products during maturation of erythroid elements by conversion of nucleotide monophosphates to diffusible nucleosides.
Paglia
D E
DE
Department of Pathology, University of California School of Medicine, Los Angeles 90024-1732.
Valentine
W N
WN
Brockway
R A
RA
Nakatani
M
M
D015720:5'-Nucleotidase; D002452:Cell Count; D003851:Deoxyribonucleases / Q000378:metabolism*; D006461:Hemolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008661:Metabolism, Inborn Errors / Q000097:blood / Q000201:enzymology / Q000235:genetics; D009708:Nucleotidases / Q000172:deficiency / Q000378:metabolism*; D012156:Reticulocytes / Q000473:pathology; D013379:Substrate Specificity
D003851:Deoxyribonucleases; D009708:Nucleotidases; D015720:5'-Nucleotidase