Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.5 (5'-nucleotidase)
 3,167 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Hepatocytes and Kupffer cells were separated from rat liver after prelabeling the Kupffer cells with colloidal iron and perfusion of the liver with digestive enzymes. The activity of several enzymes from Kupffer cells and hepatocytes was compared to validate this method of cell separation. The ratios of hepatocyte to Kupffer cell specific activities of glucose-6-phosphatase, 5'-nucleotidase, adenylate cyclase, and acid phosphatase were 20, 0.39, 0.18, and 0.078, respectively. Adenylate cyclases from hepatocytes and Kupffer cells were stimulated by fluoride ion, GTP, and catecholamines. Hepatocyte adenylate cyclase was also stimulated by glucagon, secretin, vasoactive intestinal polypeptide, and by prostaglandin E1, whereas, the Kupffer cell enzyme was completely insensitive to these hormones. The stimulation of hepatocyte adenylate cyclase by combinations of glucagon plus secretin, or glucagon plus vasoactive intestinal polypeptide, were equivalent to the sum of the individual stimulations. This suggests that the hepatocyte has specific receptors for glucagon and for vasoactive intestinal polypeptide and secretin. Prostaglandin E1 stimulation of hepatocyte adenylate cyclase was not additive to the stimulation caused by polypeptide hormones or catecholamines, nor did prostaglandin E1 decrease stimulation caused by these hormones. Although prostaglandin-sensitive adenylate cyclase was recovered with hepatocytes, 40 to 50% of the total liver prostaglandin-sensitive activity was recovered in a fraction of cell debris mixed with small cells which did not phagocytize colloidal iron.
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PMID:Stimulation of adenylate cyclase from isolated hepatocytes and Kupffer cells. 17 Dec 69

Intracellular movement of cell surface 5'-nucleotidase was studied in H4S cells, a rat hepatoma cell line. Surface labelled cells were incubated for various periods at 37 degrees C and treated with neuraminidase at 0 degrees C. Removal of sialic acid residues from glycoproteins results in a change of their isoelectric points. Analysis with isoelectric focusing was then used to distinguish between cell surface and intracellular 5'-nucleotidase. Incubation of 125I-surface-labelled cells at 37 degrees C resulted in a gradual decrease of labelled 5'-nucleotidase at the plasma membrane until, at 60 to 90 min, a steady state was reached with 52% of the label on the cell surface and 48% intracellular. Pretreatment of the cells with the weak base primaquine had no influence on this distribution while at the same time uptake of iron via the transferrin receptor was inhibited. Using immunoelectron microscopy 5'-nucleotidase was found on the cell surface, in multivesicular endosomes and the Golgi complex. Preincubation of the cells in the presence of cycloheximide caused a reduction of labelling in the Golgi complex, whereas the label in the other compartments was retained. These results lead to the conclusion that 5'-nucleotidase does not recycle through the Golgi complex and that in contrast to the transferrin receptor the recycling of 5'-nucleotidase is not inhibited by primaquine.
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PMID:Recycling of 5'-nucleotidase in a rat hepatoma cell line. 285 Jan 62

Light- and electron-microscopic histochemical procedures were used to show the distribution of the membrane-bound enzymes alkaline phosphatase (Alp), adenosine triphosphatase (ATPase), and 5'-nucleotidase (5'-nuc) in the livers of lamprey, Petromyzon marinus, throughout the life cycle. In larvae, the three enzymes are located at the biliary pole on the canalicular membranes of microvilli. At metamorphosis the enzymes become localized at all lateral cell surfaces of hepatocytes as bile canaliculi degenerate in the programmed regression of the entire biliary tree. This latter pattern of enzyme distribution persists during the parasitic adult phase but no activity is evident in individuals in the spawning migration. As the timing of the relocalization of enzymatic activity correlates well with a build-up of bile products and iron during metamorphosis, it is suggested that the lateral surface may be the new site for transport of these products.
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PMID:Relocalization of membrane enzymes accompanies biliary atresia in lamprey liver. 632 18

Thalassemic patients with iron overload are presently treated with deferoxamine or the experimental chelator deferiprone. To understand how these agents remove iron from the liver, cultured human hepatoma cells loaded with iron were previously used as a model for hepatic iron overload. The present study was undertaken to characterize further the pathways of iron transport and distribution in these cells. The activation energy for Fe2+ transport is 19 kJ/mol greater than for Fe3+, and the rate of Fe2+ transport--but not that of Fe3(+)--decreases with temperature above 25 degrees C, suggesting distinct uptake processes for different redox states of iron. Iron loading, which promotes a greater rate of Fe3+ transport, also caused a proportionally greater deposition of iron in the microsomal and cytosolic compartments and specifically lowered the activities of succinate-cytochrome c reductase and 5'-nucleotidase, representative markers of the mitochondria and plasma membrane, respectively. Both deferiprone and deferoxamine decreased total cellular iron and iron in each fraction except cytosol, indicating mobilization of iron for clearance from the cell via the cytosol. This model may be useful in characterizing the determinants of effective chelation in patients.
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PMID:Iron transport and subcellular distribution in Hep G2 hepatocarcinoma cells. 784 79

Oxidative stress and adenine nucleotide catabolism occur concomitantly in several disease states, such as cardiac ischaemia-reperfusion, and may act as synergistic determinants of tissue injury. However, the mechanisms underlying this potential interaction remain ill-defined. We examined the influence of oxidative stress on the molecular, kinetic and regulatory properties of a ubiquitous AMP-catabolizing enzyme, adenylate deaminase (AMPD) (EC 3.5.4.6). To this intent, rabbit heart AMPD and an H2O2/ascorbate/iron oxidation system were employed. Enzyme exposure to the complete oxidation system acutely impaired its catalytic activity, lowered the Vmax. by 7-fold within 5 min, and rendered the enzyme unresponsive to nucleotide effectors. Irreversible AMPD inactivation resulted within about 15 min of oxidative insult and was not prevented by free-radical scavengers. Oxidative stress did not affect the molecular mass, tetrameric nature, Km, immunoreactivity or trypsinolytic pattern of the enzyme; nor did it induce carbonyl formation, Zn2+ release from the holoenzyme or net AMPD S-thiolation. This injury pattern is inconsistent with a radical-fragmentation mechanism as the basis for the oxidative AMPD inactivation observed. Rather, the sensitivity of the enzyme to both S-thiolation and thiol alkylation and the significant (3 of 9/mol of denatured enzyme) net loss of DTNB-reactive thiols on exposure to oxidant strongly implicate the conversion of essential thiol moieties into stable higher-oxidation states in the oxidative inactivation of cardiac AMPD. The altered thiol status of the enzyme on oxidative insult may prohibit a catalytically permissible conformation and, in so doing, increase AMP availability to 5'-nucleotidase in vivo.
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PMID:Oxidative modulation and inactivation of rabbit cardiac adenylate deaminase. 788 95

In view of the profound functional and structural abnormalities shown in our previous studies in cultured, iron-loaded rat heart cells, we have examined the ability of the orally effective iron chelators dimethyl-3-hydroxypyrid-4-one (DMHP or L1) and diethyl-3-hydroxy-pyrid-4-one (DEHP or CP94) and of deferoxamine (DF) to reverse the damage caused by iron loading to heart cell organelles. At a concentration of 1.0 mmol/L, all three iron chelators were equally efficient in removing iron and restoring the activity of the thiolic sarcolemmal enzymes 5'-nucleotidase and Na,K,ATPase. However, at 0.1 mmol/L DMHP and DEHP were less effective than DF both in their iron-mobilizing effect and in promoting thiolic enzyme recovery. The superior efficiency of DF at low concentrations illustrates the advantage of the hexadentate chelating action of DF as compared with bidentate chelators such as DMHP and DEHP requiring a 3 to 1 molar ratio to iron for optimal effect. In contrast to its beneficial effect on sarcolemmal enzyme activity, iron chelation was unable to reverse the increase in beta-hexosaminidase activity caused by abnormal lysosomal fragility. Our study demonstrates for the first time that iron-induced peroxidative damage to the myocardial cell is associated with a marked loss of Na,K,ATPase activity, an enzyme with a major role in the maintenance of cellular resting potential. The timing of this damage and the restoration of Na,K,ATPase function by iron-chelating treatment suggest a cause-and-effect relationship between the observed injury to the sarcolemmal enzyme and the reversible electrophysiologic abnormalities observed in the same heart culture system in our previous studies.
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PMID:Ability of the orally effective iron chelators dimethyl- and diethyl-hydroxypyrid-4-one and of deferoxamine to restore sarcolemmal thiolic enzyme activity in iron-loaded heart cells. 816 47

The mechanism of damage to myocardial subcellular organelles was studied in iron-loaded rat myocardial cells in culture in an attempt to identify the primary target of iron's toxic effects. Lysosomes and sarcolemmal membranes were purified by fractionation of the postnuclear supernatant on a 6.7% colloidal polyvinylpyrrolidone-coated silica gradient. After 24-hour incubation with ferric ammonium citrate at a concentration of 20 micrograms/ml (0.36 mmol/L) iron, a selective depletion of polyunsaturated fatty acids was found in whole-cell homogenates, as well as in the postnuclear supernatant and sediment. Iron loading resulted in a sharp increase in the total activity of the lysosomal enzyme beta-hexosaminidase in unfractionated whole-cell homogenates, increased free enzyme activity, and loss of latent activity indicating increased lysosomal fragility. Conversely, iron loading resulted in a marked decrease in the activity of the sarcolemmal enzyme 5'-nucleotidase and a significant loss of total protein sulfhydryl group content. These studies in cultured heart cells are in agreement with previous observations indicating increased lysosomal fragility in iron-loaded hepatic and splenic tissues, attributed to increased membrane lipid peroxidation. In addition, the marked decrease in sarcolemmal 5'-nucleotidase activity and in total protein sulfhydryl group content imply that iron-induced peroxidative damage to membrane proteins may be a more important mechanism in the pathogenesis of altered myocardial function in the iron-loaded heart than formerly was recognized.
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PMID:Iron loading of cultured cardiac myocytes modifies sarcolemmal structure and increases lysosomal fragility. 842 74

To evaluate the effects of marginal zinc (Zn) deficiency on Zn absorption and metabolism, three groups of infant rhesus monkeys (n = 4/group) were fed from birth to 5 months of age either a regular infant formula (5 mg Zn/L) or a low-Zn formula (1 mg Zn/L). Since iron (Fe) intake may affect Zn absorption, the low-Zn formula was given without (1 mg Fe/L) or with Fe fortification (12 mg/L). At monthly intervals, Zn absorption and retention were assessed by gavage feeding with 65Zn and whole-body counting immediately after and on days 4, 7, and 11 after intubation. Blood samples were drawn before dosing for analyses of various potential markers of Zn status. Infants fed low-Zn formula had lower weight gain than controls; however, length growth was similar in all groups. 65Zn retention was considerably higher in both groups fed low-Zn formula (40%) than in the control group (20%), whereas plasma Zn levels were normal in all infants. Plasma metallothionein levels were generally very low and detectable in only 5 samples of 48; however, 4 of these were found in control infants. Neutrophil chemotaxis assessed at the end of the study was impaired in low-Zn infants compared to controls. In addition, low-Zn infants had increased levels of interleukin-2 at the end of the study. No differences were seen between the groups in hemoglobin levels, total white blood cells/absolute neutrophil counts, or plasma activities of 5'-nucleotidase or angiotensin converting enzyme. In conclusion, marginal Zn intake in infant rhesus monkeys resulted in increased Zn retention, which was not enough to completely compensate for the lower Zn intake. The higher level of iron fortification studied did not affect Zn retention significantly.
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PMID:Effect of infant formula zinc and iron level on zinc absorption, zinc status, and immune function in infant rhesus monkeys. 864 84

The role of iron in anthracycline toxicity was studied in rats in vivo in intact animals and in vitro in heart cell cultures. In animals treated with 8 mg/kg doxorubicin, iron loading resulted in severe weight loss and a twofold increase in rate of mortality. Studies in cultured heart cells aimed at defining the subcellular target of interaction between iron and anthracycline toxicity showed no evidence of anthracycline-induced damage to sarcolemmal thiolic enzymes represented by 5'-nucleotidase and only a limited increase in lysosomal fragility as monitored by an increase in beta-hexosaminidase activity in cell homogenates and its release into the culture medium. By contrast, doxorubicin treatment resulted in a marked inhibition of mitochondrial function as monitored by a decrease in carbon 14-labeled palmitate utilization, to 33% +/- 4% of controls, and prior iron loading resulted in a further decrease in palmitate utilization, to 18% +/- 3% of controls. Conversely, iron-chelation treatment by either deferoxamine or deferiprone (L1) eliminated the harmful effects of iron loading and resulted in a partial inhibition of doxorubicin toxicity in both normal and iron-loaded cells. Our studies represent the first demonstration in intact animals of the potentiation of anthracycline toxicity by iron overload. They also indicate that mitochondria represent an important target of combined iron-anthracycline toxicity. These observations provide new insights into the mechanism of anthracycline cardiotoxicity and may be useful in developing better strategies for tumor therapy.
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PMID:Role of iron in the potentiation of anthracycline cardiotoxicity: identification of heart cell mitochondria as a major site of iron-anthracycline interaction. 927 60

Haemophilus influenzae is a bacterium of pharmaceutical interest of which the entire genome has been sequenced. Identification of low-abundance proteins in a two-dimensional map is important for the detection of new drug targets. We applied chromatography on Polybuffer Exchanger (chromatofocusing) in order to fractionate and enrich H. influenzae proteins, possibly low-copy-number gene products, from larger volumes. Two proteins, major ferric iron-binding protein (HI0097) and 5'-nucleotidase (HI0206) were obtained in pure form and hypothetical protein HI0052 was purified to near homogeneity by this single purification step. Four other proteins, aspartate ammonia lyase (HI0534), peptidase D (HI0675), elongation factor Ts (HI0914) and 5-methyltetrahydropteroyltriglutamate methyltransferase (HI1702), were strongly enriched so that chromatography on Polybuffer Exchanger can be used as an initial step for their isolation. Approximately 125 proteins were identified in the fractions collected from the column. Seventy of these were for the first time identified after chromatography on Polybuffer Exchanger. The proteins enriched by the chromatofocusing step include both low-abundance as well as high-copy-number gene products. They do not belong to a single protein class and the majority of them are enzymes with various functions. The results include a list and a two-dimensional map of the proteins enriched by chromatofocusing. They may be useful in the search of drug targets and in the design of purification protocols for the isolation of homologous proteins from related microorganisms.
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PMID:Enrichment and purification of proteins of Haemophilus influenzae by chromatofocusing. 964 84


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