Hereditary Hemolytic Anemia with Human Erythrocyte Pyrimidine 5′-Nucleotidase Deficiency

Valentine, William N.; Fink, Kay; Paglia, Donald E.; Harris, Susan R.; Adams, William S.

doi:10.1172/jci107826

Cited by 258 publications

(148 citation statements)

References 26 publications

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“…Abbreviations: PN-I, pyrimidine 5'-nucleotidase type I; PN-II, pyrimidine nucleotidase type II; 5'-AZT-MP, 3'-azido-3'-deoxy-thymidine-5'-monophosphate; 5'-Ara-CMP, cytosine-ß-D-arabinofuranoside-5'-monophosphate; 5'-FdUMP, 5-fluoro-deoxy-uridine-5'-monophosphate; AZT, 3'-azido-3'-deoxy-thymidine; AraC, cytosine-i?-D-arabinofuranoside; 5FdUrd, 5-fluoro-2'-deoxy-uridine; DTT, dithiothreitol; p-CMB, /)-chloromercuribenzoate; DTNB, 5,5'-dithiobis(2'-nitrobenzoic acid) Such a condition determines a marked erythrocyte accumulation of pyrimidine nucleotides, detectable both by spectrophotometrically [1] and by a HPLC-based assay [4], and is accompanied by a distinctive basophilic stippling of erythrocytes upon Wright staining of blood smears [1]. Furthermore it has been shown [3,5,6] that affected patients, although defective in erythrocyte 5'-nucleotidase activity towards UMP, CMP and dCMP, conserved normal activity toward dUMP and dTMP.…”

Section: Introductionmentioning

confidence: 99%

Pyrimidine nucleotidases from human erythrocyte possess phosphotransferase activities specific for pyrimidine nucleotides

et al. 1997

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Two cytoplasmic forms of pyrimidine nucleotidase (PN-I and PN-II) have been purified from human erythrocytes to apparent homogeneity and partially characterized. They preferentially hydrolyse pyrimidine 5'-monophosphates and 3'-monophosphates respectively. PN-I and PN-II operate as interconverting activities, capable of transferring the phosphate from the pyrimidine nucleoside monophosphate donor(s) to various nucleoside acceptors, including important drugs like 3'-azido-3'-deoxy-thymidine (AZT), cytosine-ß-D-arabinofuranoside (AraC) and 5-fluoro-2'-deoxy-uridine (5FdUrd), pyrimidine analogues widely used in chemotherapy. Kinetic analysis showed linear behaviour for both PN-I and PN-II. PN-I phosphotransferase activity revealed higher affinity for oxynucleosides with respect to deoxy-nucleosides, whereas the contrary seems to be true for PN-II. These results show for the first time that soluble pyrimidine nucleotidases are endowed with pyrimidine-specific phosphotransferase activity.

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Section: Introductionmentioning

confidence: 99%

Pyrimidine nucleotidases from human erythrocyte possess phosphotransferase activities specific for pyrimidine nucleotides

et al. 1997

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“…In addition, pyrimidine nucleotidases also possess phosphotransferase activity between pyrimidine mononucleotides and pyrimidine nucleosides (3,4). Two different subtypes, hP5N-1 2 (UniProt code Q9H0P0) and hPN-2, which differ in their substrate specificities, were identified in human erythrocytes (5)(6)(7). Subtype hP5N-1 preferentially dephosphorylates 5Ј-UMP, 5Ј-CMP, and at a lower rate also 5Ј-dCMP, 5Ј-dTMP, and 5Ј-dUMP, whereas hPN-2 preferentially dephosphorylates 3Ј-dTMP, 3Ј-dUMP, and 3Ј-UMP but shows no activity against 5Ј-CMP and 5Ј-dCMP (3,4,8).…”

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confidence: 99%

Structure of Pyrimidine 5′-Nucleotidase Type 1

Bitto

Bingman

Wesenberg

et al. 2006

Journal of Biological Chemistry

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Eukaryotic pyrimidine 5-nucleotidase type 1 (P5N-1) catalyzes dephosphorylation of pyrimidine 5-mononucleotides. Deficiency of P5N-1 activity in red blood cells results in nonspherocytic hemolytic anemia. The enzyme deficiency is either familial or can be acquired through lead poisoning. We present the crystal structure of mouse P5N-1 refined to 2.35 Å resolution. The mouse P5N-1 has a 92% sequence identity to its human counterpart. The structure revealed that P5N-1 adopts a fold similar to enzymes of the haloacid dehydrogenase superfamily. The active site of this enzyme is structurally highly similar to those of phosphoserine phosphatases. We propose a catalytic mechanism for P5N-1 that is also similar to that of phosphoserine phosphatases and provide experimental evidence for the mechanism in the form of structures of several reaction cycle states, including: 1) P5N-1 with bound Mg(II) at 2.25 Å , 2) phosphoenzyme intermediate analog at 2.30 Å , 3) product-transition complex analog at 2.35 Å , and 4) product complex at 2.1 Å resolution with phosphate bound in the active site. Furthermore the structure of Pb(II)-inhibited P5N-1 (at 2.35 Å ) revealed that Pb(II) binds within the active site in a way that compromises function of the cationic cavity, which is required for the recognition and binding of the phosphate group of nucleotides.Eukaryotic pyrimidine nucleotidases are enzymes involved in catabolism of RNA and in the pyrimidine salvage pathway (1, 2). These enzymes catalyze dephosphorylation of pyrimidine mononucleotides to their respective nucleosides. In addition, pyrimidine nucleotidases also possess phosphotransferase activity between pyrimidine mononucleotides and pyrimidine nucleosides (3, 4). Two different subtypes, hP5N-1 2 (UniProt code Q9H0P0) and hPN-2, which differ in their substrate specificities, were identified in human erythrocytes (5-7). Subtype hP5N-1 preferentially dephosphorylates 5Ј-UMP, 5Ј-CMP, and at a lower rate also 5Ј-dCMP, 5Ј-dTMP, and 5Ј-dUMP, whereas hPN-2 preferentially dephosphorylates 3Ј-dTMP, 3Ј-dUMP, and 3Ј-UMP but shows no activity against 5Ј-CMP and 5Ј-dCMP (3,4,8). Both enzymes are essentially inactive against purine nucleotides. This specificity of hPNs is unique among 5Ј-nucleotidases (EC 3.1.3.5), which are typically not discriminatory to the identity of the nucleotide base (9). The specificity of hPNs is crucial in erythrocytes for preservation of the internal pool of ATP/GTP (6). Two types of hP5N-1 deficiency are known: hereditary and acquired. Hereditary deficiency is associated with nonspherocytic hemolytic anemia (NHA) (2, 7, 10). Acquired deficiency results from inhibition of hP5N-1 by Pb(II) during lead poisoning, and its severity is related to the blood levels of Pb(II) (11)(12)(13)(14). Both NHA and severe lead poisoning (Ͼ80 g/dl Pb(II) in blood) have the same clinical manifestation: erythrocytes in blood smears of affected patients show a distinct basophilic stippling upon Wright's staining (7,14,15). At the molecular level, NHA is characterized by signifi...

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“…Our recent study [7] demonstrated that the serum from dogs infected with B. gibsoni dose-dependently retarded the maturation of canine reticulocytes and decreased the activity of erythrocyte 5'-nucleotidase, which is involved in the morphological maturation of reticulocytes by contributing to the degradation of reticulocyte RNA in ribosomes [21]. The in vitro multiplication of B. gibsoni also induced a significant decrease of this enzyme activity [7].…”

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confidence: 94%

Inhibitory Effect of Pyrimidine and Purine Nucleotides on the Multiplication of <i>Babesia gibsoni</i>: Possible Cause of Low Parasitemia and Simultaneous Reticulocytosis in Canine Babesiosis

Hossain

Yamato

Yamasaki

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. The present study was conducted to determine the cause of low parasitemia and simultaneous reticulocytosis in canine babesiosis. The parasitemia was significantly decreased in in vitro cultures of Babesia gibsoni by the pretreatment of host canine erythrocytes with lead acetate, which is a specific inhibitor of pyrimidine 5'-nucleotidase subclass I (P5N-I). The serum from dogs chronically infected with B. gibsoni did not decrease the activities of hexokinase, glucose-6-phosphate dehydrogenase or 6-phosphogluconate dehydrogenase in canine reticulocytes, although it was previously reported that this serum had inhibitory effects on both the maturation of reticulocytes and the canine P5N-I and purine-specific 5'-nucleotidase activities. Furthermore, the in vitro multiplication of B. gibsoni was significantly inhibited by pyrimidine nucleotides such as cytidine 5'-monophosphate (5'-CMP), which is preferentially catalyzed by P5N-I and also inhibits the morphological maturation of canine reticulocytes. Purine nucleotides such as inosine 5'-monophospha te (5'-IMP) also had an inhibitory effect on the multiplication of this parasite. These results suggest that nucleotides such as 5'-CMP and 5'-IMP might accumulate in young erythrocytes and/or serum in dogs infected with B. gibsoni as a result of the decreased activity of erythrocyte 5'-nucleotidase, and the accumulation of these nucleotides might inhibit the multiplication of this parasite and simultaneously retard the maturation of reticulocytes. The results obtained from the in vitro examinations in the present study may partially clarify the relationship between low parasitemia and simultaneous reticulocytosis in vivo in canine babesiosis. KEY WORDS: Babesia gibsoni, erythrocyte 5'-nucleotidase, purine nucleotide, pyrimidine nucleotide, reticulocyte.

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Hereditary Hemolytic Anemia with Human Erythrocyte Pyrimidine 5′-Nucleotidase Deficiency

Cited by 258 publications

References 26 publications

Pyrimidine nucleotidases from human erythrocyte possess phosphotransferase activities specific for pyrimidine nucleotides

Pyrimidine nucleotidases from human erythrocyte possess phosphotransferase activities specific for pyrimidine nucleotides

Structure of Pyrimidine 5′-Nucleotidase Type 1

Inhibitory Effect of Pyrimidine and Purine Nucleotides on the Multiplication of <i>Babesia gibsoni</i>: Possible Cause of Low Parasitemia and Simultaneous Reticulocytosis in Canine Babesiosis

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