Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “NUCLEOTIDASES”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 829 records · Page 46Linked to original sources
PubMed1998

Ecto-nucleotidases terminate purinergic signalling in the cochlear endolymphatic compartment.

There is strong evidence for a purinergic signalling system in the inner ear which regulates auditory sensitivity. This study describes the terminating mechanism for purinergic signalling in the cochlear endolymphatic compartment via ecto-nucleotidases. Exogenous ATP was introduced into the scala media (SM) of the isolated, perfused guinea-pig cochlea, and the effluent was assayed for the adenine nucleotide metabolites by reverse-phase HPLC. Tissue viability was confirmed by fluorescence imaging of cochlear tissues. Extracellular ATP degradation to adenosine was Ca2+/Mg2+ dependent, and was not affected by inhibitors of intracellular ATPases and non-specific alkaline phosphatase. High azide concentration (5 mM) and suramin produced an inhibitory effect on ATP hydrolysis, consistent with inhibition of E-type ATPase activity. The Vmax of ATP hydrolysis (2564 mumol min-1 SM-1) was indicative of high ecto-ATPase activity. Our results support the role of ecto-nucleotidases as a principal mechanism for termination of purinergic signalling within SM, a compartment of the cochlea showing considerable P2X receptor expression.

Adenosine↗
PubMed1986

Decreased adenosine deaminase (ADA) and 5'nucleotidase (5NT) activity in peripheral blood T cells in Hodgkin disease.

The purine metabolic enzymes adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and 5'nucleotidase (5NT) play an important role in normal lymphocyte differentiation. Abnormal levels of one or all of these enzymes have been associated with immunodeficiency diseases and lymphoproliferative disorders. ADA, PNP, and 5NT activity was measured in peripheral blood T cells from 24 patients with Hodgkin disease (HD) (12 in complete remission and 12 with active disease) to determine whether an association existed between enzyme abnormalities and the decreased cellular immune function previously described in this disorder. HD patients had a significantly decreased absolute lymphocyte count (1,618 +/- 1107/mm3; mean +/- SD) compared to controls (2,320 +/- 980; p less than .001). ADA, PNP, and NT activity was assessed in lymphocyte extracts by measuring the conversion of radiolabeled substrates to products over time. ADA activity expressed as mean +/- SEM nanomoles/10(6) lymphocytes/hr was significantly decreased in T cells from HD patients (84.6 +/- 7.5) compared to controls (128 +/- 12.3; p less than 0.025). Likewise, 5NT was significantly decreased in HD patients (12.7 +/- 1.3) compared to controls (24.0 +/- 3.6; p less than .005). There was not a significant difference in PNP activity between both groups. Low 5NT activity was present irrespective of whether patients had active disease (12.1 +/- 1.5) or were in unmaintained complete remission (14.5 +/- 2.4). These findings suggest that biochemical abnormalities may be responsible for or related to the persistent abnormalities in T-cell function noted throughout the clinical course of HD.

5'-Nucleotidase↗
PubMed1987

Erythrocyte-oxidized glutathione transport in pyrimidine 5'-nucleotidase deficiency.

The oxidized form of glutathione transport was studied in human erythrocytes in pyrimidine 5'-nucleotidase (P5N) deficiency, a disorder in which the amounts of CTP and UTP in the erythrocytes are elevated. The inhibition of ATP-requiring oxidized glutathione (GSSG) transport by CTP and UTP is believed to play a role in elevating the levels of the reduced form of glutathione (GSH) in the erythrocytes of patients with P5N deficiency. The current investigation was undertaken to determine if GSSG transport actually decreases in the erythrocytes of such patients. Erythrocytes from a 17-year-old patient and a 13-year-old patient with P5N deficiency hemolytic anemia and from ten normal subjects were used as materials for the experiment. Erythrocytes, which had been previously incubated with [3H]glycine, were incubated at 37 degrees C, and the rate of [3H]GSSG transported by the cells was estimated. The velocity of GSSG transport out of the erythrocytes was quite low in the patients, 3.17-3.65 nmol GSSG/ml erythrocytes/hr at 37 degrees C in one case, and 3.30 nmol GSSG/ml erythrocytes/hr in the other case, vs that in the normal controls (6.00 +/- 0.80 nmol GSSG/ml erythrocytes/hr; mean +/- SD). The activity of gamma-glutamylcysteine synthetase and glutathione synthetase did not decrease in the patients. Decreased transport activity of GSSG in addition to a normal synthesis rate for GSH may explain the increased concentration of erythrocyte GSH in P5N deficiency.

5'-Nucleotidase↗
PubMed1986

Ecto-5'nucleotidase activity in B and T lymphocytes and monocytes of patients with multiple sclerosis.

Ecto-5'nucleotidase activity in the plasma membrane of B and T lymphocytes and cultured and noncultured monocytes from patients with multiple sclerosis and from age- and sex-matched control subjects was determined by radioisotopic assay. The activity on lymphocytes and noncultured monocytes was normal, but the activity on cultured monocytes was found to be higher than normal in 70% of patients with active disease and lower than normal in 85% of patients with chronic inactive disease. Immunofluorescence staining with monoclonal antibodies, carried out in parallel with each assay, did not show any phenotypic differences between each cell population and its control.

5'-Nucleotidase↗
PubMed1988

Reduction of monocyte 5'nucleotidase activity by gamma-interferon in multiple sclerosis and autoimmune diseases.

Ecto-5'nucleotidase (5'NT) activity in the plasma membrane of peripheral blood monocytes from patients with multiple sclerosis (MS), rheumatoid arthritis (RA), myasthenia gravis (MG), motor neuron disease (MND), and from normal control subjects of similar age was determined by radioisotopic assay. The activity in unstimulated monocytes cultured for 24 and 48 hours was found to be higher than normal in 56% of patients with active relapsing MS, 29% of patients with RA, and 7% of patients in the MG/MND group. While the enzyme activity was reduced after cultivation of the cells with recombinant interferon-gamma (gamma-IFN) in all of the groups studied, the percentage reduction was significantly greater in monocytes from patients with active relapsing MS who were in relapse at the time of sampling (p = 0.03). HLA-DR expression was monitored using immunofluorescence staining with monoclonal antibody and showed that similar numbers of monocytes in all patient and control groups expressed this antigen. Thus, while high monocyte 5'NT activity was found principally in patients with active relapsing MS and in some patients with RA, the monocytes from patients with MS were, in addition, exquisitely sensitive to stimulation with gamma-IFN.

5'-Nucleotidase↗
PubMed1984

Redistribution of ecto-5'-nucleotidase during phagocytosis by guinea pig polymorphonuclear leukocytes.

We demonstrate that 5'-nucleotidase (5'NT), an ectoenzyme of guinea pig polymorphonuclear leukocytes, is largely excluded from phagosomal membrane, rather than internalized randomly during phagocytosis of heat-killed bacteria, latex microbeads, or zymosan particles. Cells were fixed in 0.25% glutaraldehyde (pH 6.3) at 4 degrees C for 10 min and incubated in a cytochemical medium for the demonstration of 5'NT. In the nonphagocytosing cells, 5'NT was evenly distributed on the external side of the plasma membrane. In cells phagocytosing bacteria, 5'NT appeared to be cleared from the nascent phagosomal membrane; after 5 min of phagocytosis, most of the phagocytic vacuoles containing bacteria, latex, or zymosan particles were devoid of reaction product. When phagosomes containing latex particles were isolated and biochemically assayed, they contained less than 3% of the total cellular 5'NT activity even after 60 min of phagocytosis, and at that time the total cellular 5'NT activity had not declined. When the diazonium salt of sulfanilic acid (DASA), a nonpermeable ectoenzyme inhibitor, was used to determine the distribution of extracellular and intracellular 5'NT activity, no increase in DASA-insensitive intracellular 5'NT was found after phagocytosis of latex or opsonized zymosan. Cytochemical and biochemical evidence led us to conclude that 5'NT is excluded from phagosomal membrane, and that the exclusion is due to redistribution rather than to inactivation by granule enzymes.

5'-Nucleotidase↗
PubMed1985

Expression of 5'nucleotidase activity and wheat-germ agglutinin binding sites in mononuclear phagocytes from bone marrow cultures.

The question as to whether the various types of mononuclear phagocyte found in bone marrow cultures and recognized by specific peroxidatic (PO) activity patterns differ in the expression of binding sites for the lectin wheat-germ agglutinin (WGA) and the activity of the ectoenzyme 5'nucleotidase (5'N) was investigated. Monoblasts, promonocytes, monocytes, and/or exudate macrophages, and exudate-resident macrophages generally showed a high level of WGA binding, and a considerably lower level was found in the PO-negative cells and in resident macrophages. 5'N activity was absent in monoblasts, promonocytes, and in the great majority of the monocytes and/or exudate macrophages, but was demonstrable in exudate-resident macrophages and resident macrophages, as well as in PO-negative macrophages after 4 days of culture. On the basis of the successive occurrence of the above-mentioned phenotypes in cultures, the possibility that this diversity in WGA binding and 5'N activity is related to modulation during cell differentiation is discussed. The present findings led to the conclusion that the PO-negative macrophages, whose origin was previously not entirely certain, are precursors of resident macrophages.

5'-Nucleotidase↗
PubMed1988

Induction of adenosine deaminase and 5' nucleotidase activity in cultured human blood monocytes and monocytic leukemia (THP-1) cells by differentiating agents.

The in vitro effect of short-term culture as well as the effect of retinol (ROH), retinoic acid (RA), muramyl dipeptide [( Abu']MDP), lipopolysaccharide (LPS), and gamma interferon (IFN-gamma) on the induction of the purine metabolic enzymes, adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and 5'nucleotidase (5NT) in human peripheral blood monocytes (HPBM) was examined. HPBM isolated by centrifugal elutriation were cultured for up to 96 h. Following an initial time lag of 24 h, mean ADA activity from seven separate experiments as measured in nmoles/10(6) cells/h increased from a baseline of 31.3 +/- 9.3 to 57.8 +/- 16.4 (P less than 0.005) at 72 h and to 72 +/- 21.5 (P less than .025) by 96 h. 5NT activity increased from a baseline of 2.2 +/- 0.9 to a maximum of 44 +/- 10.1 by 72 h and then declined to 29 +/- 18 (P less than 0.005) by 96 h, while no significant change in PNP activity was observed. HPBM incubated for 3 d with optimal concentrations of LPS, RA, and IFN-gamma had increases in ADA and 5NT activity ranging from three- to 10-fold compared to HPBM cultured in media alone, whereas no effect was observed with ROH and [Abu']MDP. RA, but not ROH, significantly enhanced ADA activity in a monocytic leukemia cell (THP-1) line. Addition of RA or the tumor promoter, phorbol 12-myristic 13-acetate (PMA), to HPBM or THP-1 cells resulted in significant increases in 5NT activity with opposite effects on ADA activity. These findings suggest that the biological mechanisms associated with differentiation in normal and malignant monocytes seem to be related and that the sequence and degree to which the various differentiation agents induce the enzyme elevations are also related to the mechanisms of activation/differentiation.

5'-Nucleotidase↗
PubMed1984

Investigations on myelinogenesis in vitro: regulation of 5'-nucleotidase activity by thyroid hormone in cultures of dissociated cells from embryonic mouse brain.

The developmental pattern of the myelin-associated 5'-nucleotidase and its regulation by L-3,3',5,-triiodothyronine (T3) have been demonstrated in a culture system of cells dissociated from embryonic mouse brain. Hypothyroid calf serum containing low levels of T3 (31 ng/100 ml), and thyroxine, T4 (less than 1 microgram/ml), was used in the culture medium in place of normal calf serum (T3, 103 ng/100ml; T4, 5.7 micrograms/ml) to render the cultures responsive to exogenously added T3. By means of T3 supplementation, the lower levels of enzyme activity observed in the cultures grown in the presence of hypothyroid calf-serum containing medium could be restored to a considerable extent although not completely to normal values. Half-maximal stimulatory effect was obtained at 3.9 X 10(-8)M T3 concentration. Among the various substrates tested, 5'-AMP, 5'-UMP and 5'-CMP were equally good, while 5'-GMP yielded approximately half the activity.

5'-Nucleotidase↗
PubMed1983

A histochemical study of variations in the localization of 5'-nucleotidase activity in the acinar cell of the rat exocrine pancreas over the twenty-four hour period.

The circadian variation of 5'-nucleotidase (AMPase) activity was studied in rat pancreatic exocrine cells. The localization of this enzyme, often associated with the plasmalemma, was studied by ultracytochemical methods at six time points over the 24-h period. The localization of AMPase activity exhibited a clear-cut circadian variation. During the light span strong activity was observed on the luminal plasmalemma, negative or weak activity on the baso-lateral plasmalemma and clearly visible activity on intracellular structures such as cytoplasmic vacuoles (fragmentation-like vesicles), dilated rims of the Golgi cisternae (or cisternal ends of the Golgi stacks), condensing vacuoles and lysosomal bodies. During the dark span the activity was detectable only on the baso-lateral plasmalemma. The fact that AMPase activity could not be found on the luminal plasmalemma during the dark span suggests that the luminal membranes may be replaced by the membranes of secretory granules, which do not display AMPase activity. The intracellular localization of AMPase activity during the light span, especially at 08.00 h, includes all cytoplasmic compartments which have hitherto been associated with the intracellular pathway for membrane retrieval from the plasmalemma. Moreover, the appearance of the activity in the dilated rims of the Golgi stack and condensing vacuoles indicates that these compartments may constitute a functional unit.

5'-Nucleotidase↗
PubMed1981

Localization of 5'-nucleotidase activity in the parotid acinar cells of a rat treated with isoproterenol.

Cytochemical localization of 5'-nucleotidase (AMPase) has been investigated in the parotid acinar cells of rats at various stages of exocytic secretion induced by an administration of isoproterenol (IPR). In the resting stage, the acinar cells show AMPase activity located on the baso-lateral and luminal plasmalemma, and in the earliest secretory stage the luminal plasma membranes are devoid of the enzymatic activity. However, these particular regions exhibit AMPase activity during the advanced stages of secretion, and the AMPase positive membranes become absorbed into the cytoplasm by endocytic activity. The absorbed membrane components then seem to be degraded by the action of lysosomes. The intracellular fate of the endocytic vacuoles has been examined by the aid of ferritin particles introduced retrogradely through ductal lumina. Ferritin containing vacuoles are distributed in the cytoplasm, and these droplets change into secondary lysosomes. No tracer particles are recognized in the internal space of the Golgi lamella and its associated vesicles. The results suggested that in the exocytic secretion of parotid acinar cells, AMPase originating from plasma membrane intermingles with the membranes derived from secretion granules, and is translocated into cytoplasm by an endocytic mechanism. The internalized membrane components are, at least partly, degraded by lysosome action.

5'-Nucleotidase↗
PubMed1988

Assignment of ecto-5'-nucleotidase to human chromosome 6.

Ecto-5'-nucleotidase activity (5NT) was measured on whole cells of 26 human x Chinese hamster hybrids. Concordance analysis showed 100% correlation between enzyme activity and inheritance of human chromosome 6. This observation was confirmed by a segregation analysis in which cells of a hybrid containing chromosome 6 were stained by indirect immunofluorescence for HLA Class 1 antigen and sorted by a fluorescence-activated cell sorter (FACS). Cells in the HLA- compartment were cloned and expression of HLA and 5NT was determined. Of nine clones, three were HLA-, 5NT- and six were HLA+, 5NT+, supporting the linkage of 5NT to chromosome 6.

5'-Nucleotidase↗
PubMed1981

Relationship between lead concentration in blood and the activities of erythrocyte pyrimidine 5'-nucleotidase and delta-aminolevulinate dehydratase in the mice exposed to lead.

The relationship between the activities of both pyrimidine 5'-nucleotidase (Py5N) and delta-aminolevulinate dehydratase (ALA-D) in erythrocytes and the concentration of lead in blood was investigated in the mice which were given ad libitum a drinking water containing lead of 10 to 500 ppm, for 30 days. From these results, it was demonstrated that the erythrocyte Py5N activity is inhibited by lead and its activity level is negatively correlated with the concentration of lead in blood (r = -0.78). In addition, it was suggested that the erythrocyte Py5N activity is a better indicator in the exposure to relatively high lead concentration while the ALA-D is a more sensitive indicator for evaluating the lead exposure of low and moderate levels.

5'-Nucleotidase↗
PubMed1984

Lack of correlation between two markers of lymphocyte differentiation: 5' nucleotidase activity and T lymphocyte colony formation.

Ecto 5' nucleotidase (5' NT) activity and T lymphocyte colony formation (TLCF) are both reputed to be markers for lymphocyte maturation. In order to determine whether these two expressions of lymphocyte activity are related, we compared 5' NT activity with the TLC forming capacity of mononuclear cells from three study groups: normal adults, cord blood, and patients with immunodeficiencies. Despite individual examples of correlation between these two measurements, there was poor overall correlation in any of the groups studied. Although both measurements may reflect maturation of certain cellular activities, these are unlikely to be related.

5'-Nucleotidase↗
PubMed1983

Independent expression of the surface markers 5'-nucleotidase and cALLA on leukemic cells.

High levels of the ectozyme 5'-nucleotidase (5'-N) and the common ALL-antigen (cALLA) are coexpressed on leukemic blast cells in common ALL, in the lymphoid blast crisis of CML and also on the lymphoblastoid cell-line Nalm-1. Clinically this coexpression can help to subclassify leukemias and may be of diagnostic and prognostic significance. In an attempt to study the mechanism underlying this simultaneous expression plasmamembrane subfractionation was undertaken on Nalm-1. When membrane-shedding from intact cells is induced by sublytic concentrations of the lysophosphatidyl-choline analogue ET-12-H, membrane subfractions are obtained which contain 30-40% of total cellular 5'-N, which is most of the enzyme carried on the cell surface, in a highly enriched form. Under these conditions only a very low release of intracellular enzymes is observed. On the other hand cALLA is not accumulated in these membrane fractions to any appreciable extent. The predominant part of this antigen is still on the intact cells remaining after the shedding procedure. It is concluded that the simultaneous expression of 5'-N and cALLA on Nalm-1 and leukemic blasts is not regulated by a physical association or a close neighborhood of these antigens on the membrane level.

5'-Nucleotidase↗
PubMed1989

Patterns of adenosine deaminase, ecto-5'-nucleotidase, poly(A)polymerase and surface light chain expression in chronic lymphocytic leukemias.

The levels of activity of three enzymes have been measured in the circulating malignant lymphocytes of 47 patients with B chronic lymphocytic leukemia (CLL). These were the purine degradative enzymes, adenosine deaminase (ADA) and ecto-5'-nucleotidase (5'NT) and the enzyme responsible for the polyadenylation of mRNA, poly(A) polymerase. The patterns of activity of the above enzymes and the expression of surface immunoglobulin light chains were examined. A heterogeneity in the specific activity of the enzymes was observed which could not be attributed to variations of the percentage of B lymphocytes. A positive correlation was found between ADA and poly(A)polymerase activity (r = 0.383, p less than 0.01). Furthermore, the expression of immunoglobulin light chain phenotype was inversely related to 5'NT specific activity; CLL cases in which less than 20% of the cells expressed lambda chain phenotype, presented 5'NT specific activity of 16.7 +/- 3.3 (S.E.) nmol/h/10(6) cells, whereas in CLL cases with more than 20% of the cells expressing this phenotype the enzyme specific activity was 4.8 +/- 1.6 (S.E.) nmol/h/10(6) cells (p less than 0.02). These findings suggest that the simultaneous determination of enzymatic activities and immunological markers, might be useful in defining subsets in CLL and the subsequent clinical treatment.

5'-Nucleotidase↗
PubMed1989

Effect of lead exposure on some biological indices related to porphyrin metabolism and the activity of erythrocyte pyrimidine 5'-nucleotidase in the mice.

The urinary excretion of delta-aminolevulinic acid (ALA) and coproporphyrin (CP) isomers (I and III) was investigated in mice exposed to lead in drinking water (200 and 500 ppm) for 14 or 30 days. Furthermore, the inhibition of pyrimidine 5'-nucleotidase (P5N) and delta-aminolevulinic acid dehydratase (ALAD) was studied in the same mice. In the lead-exposed mice, urinary excretion of ALA was significantly elevated (12-times control), while that of CP remained unchanged. This discrepancy on the urinary excretion may be of interest. The P5N activity in both erythrocytes and bone marrow cells was significantly inhibited by exposure to lead. However, the degree of inhibition was greater in the erythrocytes (45% inhibition) than in the bone marrow cells (25% inhibition). The inhibition of ALAD was much higher in erythrocytes (90% inhibition) than in liver (20-40% inhibition). The increase in erythrocyte protoporphyrin and the decrease in both hemoglobin and hematocrit were not observed in mice exposed to lead under the above conditions.

5'-Nucleotidase↗
PubMed1988

Relationship between inhibition of erythrocyte pyrimidine 5'-nucleotidase activity and biological response for porphyrin metabolism in workers occupationally exposed to lead.

A rapid determination of erythrocyte pyrimidine 5'-nucleotidase (P5N) activity in lead workers was carried out using a high-performance liquid chromatograph (HPLC). The P5N activity had a good negative correlation with the concentration of lead in blood (PbB) ranging from 16 to 96 micrograms/dl (r = -0.82, n = 77). Further, the P5N was compared with other biological parameters: erythrocyte delta-aminolevulinic acid dehydratase (ALAD) activity, erythrocyte protoporphyrin (PROTO), urinary delta-aminolevulinic acid (ALA) and urinary coproporphyrin (COPRO). The correlation coefficients between P5N and ALAD, log PROTO, log ALA, and log COPRO were 0.59, -0.72, -0.65, and -0.61, respectively. On the other hand, the normal value of P5N obtained from 72 healthy subjects was 11.9 +/- 2.1 units; mumol uridine/h/g Hb (mean +/- SD), indicating that the lower limit of 95% confidence interval for normal P5N was about 8 units. When P5N was cut off at less than or equal to 8 units in 77 lead workers, the validity (sensitivity + specificity) for PbB greater than or equal to 40 micrograms/dl, PbB greater than or equal to 60 micrograms/dl, erythrocyte PROTO greater than or equal to 150 micrograms/dl RBC, urinary ALA greater than or equal to 6 mg/l, and urinary COPRO greater than or equal to 150 micrograms/l was 1.66, 1.76, 1.57, 1.68, and 1.60, respectively. From these results, it was confirmed that the erythrocyte P5N test is suitable for the biological monitoring of exposure to lead in a wide range, and its activity is useful in predicting the disturbance of porphyrin metabolism induced by lead.

5'-Nucleotidase↗