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)

The controversial subject of mitochondrial 5'-nucleotidase in the liver was studied employing density gradient fractionation combined with a method for analyzing the distribution profiles of marker enzymes based on multiple regression analysis. Triton WR-1339 was used to improve the separation of mitochondria from lysosomes by the gradient centrifugation technique. Adenosine production was examined further using acetate to increase intramitochondrial AMP, and thus adenosine production, in incubations with gradient centrifugation-purified mitochondria. Distribution analysis of the crude homogenate showed that 5'-nucleotidase activity exists in the mitochondrial fraction. To increase the resolution of this approach with respect to mitochondria, a crude mitochondrial fraction was also studied. In this case the relative mitochondrial activity decreased but 5'-nucleotidase activity was still clearly detectable. The mitochondrial 5'-nucleotidase exhibited a Km of 94 microM and a Vmax of 31 nmol/min per mg protein for AMP. The kinetic data for the Mg2+, ATP, ADP and AOPCP sensitivity of the enzyme showed that it differs from the plasma membrane, lysosome and cytosol 5'-nucleotidases. AOPCP was only a moderate inhibitor, and ATP was a more potent inhibitor than ADP at a 1 mM concentration. The enzyme also showed a requirement of Mg2+. Acetate caused the conversion of intramitochondrial adenylates to AMP and the formation of adenosine. Adenosine concentration increased in the extramitochondrial space in a time-dependent manner, but only trace amounts of nucleotides were detected. The data show that 5'-nucleotidase activity producing adenosine exists in rat liver mitochondria and a concentration-dependent adenosine output from mitochondria by diffusion or facilitated diffusion is also suggested.
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PMID:5'-Nucleotidase activity and adenosine production in rat liver mitochondria. 155 Aug 32

In view of its vasodilatory effect on the coronary circulation (probably mediated by adenosine) and its metabolic compartmentalization (intramitochondrial activation to form acetyl-CoA), the metabolic effects of acetate were studied in isolated rat heart mitochondria. Acetate caused conversion of adenylates to AMP and the formation of adenosine. Adenylate efflux was inhibited by carboxyatractyloside but not by N-ethylmaleimide. The intramitochondrial accumulation of AMP was enhanced by carboxyatractyloside during acetate metabolism and the formation of extramitochondrial adenosine inhibited. A carboxyatractyloside-sensitive unidirectional AMP influx with a Km of 50 microM and Vmax of 11 nmol/min per mg mitochondrial protein was also observed. The mitochondrial adenosine content was high and constant during the experiments. The steep apparent concentration gradient of adenosine indicates that most of the mitochondrial adenosine is tightly bound to protein. Adenosine formation was proportional to the extramitochondrial AMP concentration, showing that the 5'-nucleotidase activity of cardiac mitochondrial preparations is extramitochondrial in origin. The data suggest that the mitochondrial ATP/ADP carrier is capable of transporting AMP and that intramitochondrial AMP is recycled during acetate metabolism in the myocardium partially by means of the ATP/ADP translocator, leading to an increase in extramitochondrial AMP and adenosine formation.
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PMID:Adenine nucleotide transport and adenosine production in isolated rat heart mitochondria during acetate metabolism. 254 56

Immunohistochemical techniques have been used to localize clotting factor XIII subunit A in human reactive lymphoid follicles. The follicular dendritic reticulum cells (DRCs) were identified by the monoclonal antibodies R4/23 and OKB-7 as well as by their 5'-nucleotidase positivity. Follicular histiocytic reticulum cells (HRCs) were demonstrated by their acid phosphatase and non-specific esterase reactions. Capillaries were selectively visualized by adenosine triphosphatase. The immunohistochemical demonstration of F-XIIIa was preferably carried out in combination with one or two of the above marker techniques, on the same cryostat section. The subunit A of factor XIII is present in follicular DRCs. Their selective immunohistochemical demonstration with antibody against F-XIIIa requires formaldehyde fixation of cryostat sections. Similar fixation, however, is inappropriate for the demonstration of F-XIIIa reactivity of DRCs in paraffin sections. For this purpose, acetic acid-formalin fixation is useful. Follicular HRCs are consistently negative for F-XIIIa, contrary to the F-XIIIa positivity of sinusoidal and interfollicular HRCs. Developmental and functional implications of F-XIIIa reactivity in DRCs and HRCs are suggested.
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PMID:Selective visualization of human dendritic reticulum cells in reactive lymphoid follicles by the immunohistochemical demonstration of the subunit A of factor XIII (F-XIIIa). 288 67

The formation of N tau-ribosylhistidine (His-R), a novel histidine derivative found in the urine of histidinemic patients, was studied. A most possible synthetic pathway catalyzed by imidazole acetic acid (ImAA) phosphoribosyltransferase was not substantiated, because p.o. administration to humans and rats of aspirin, an inhibitor of the enzyme, did not change the urinary excretion of His-R, whereas aspirin decreased the excretion of ImAA-R with concomitant increase in that of ImAA. His-R was produced on incubation of a rat liver homogenate or its membrane fraction with histidine, NAD(P)+ and MgCl2, but not with only histidine or NAD(P)+. Nicotinamide inhibited the formation of His-R. Thus the enzymes responsible for the formation of His-R were suggested to be NAD(P)+ nucleosidase, nucleotide pyrophosphatase and 5'-nucleotidase.
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PMID:Formation of N tau-ribosylhistidine, a novel histidine derivative found in the urine in histidinemia, from histidine and NAD(P)+ catalyzed by an NAD(P)+ glycohydrolase system. 299 72

Purified pig pancreatic zymogen granules were subjected to free flow electrophoresis (FFE) in an acetate buffer system (acetic acid/NaOH, pH 5.5) to detect the presence or absence of more than one population or zymogen granules. Pig pancreatic zymogen granules were purified by differential and density gradient centrifugation and subjected to FFE. Fractions were analyzed for protein, alpha-amylase (EC 3.2.1.1) and 5'-nucleotidase (EC 3.1.3.5) as marker enzymes for zymogen granule content and membranes, respectively. Only one distinct peak, with coincident alpha-amylase and 5'-nucleotidase activity, and most protein was detected, which reflects the presence of a single population of intact zymogen granules. This was confirmed by electron microscopy. When the granules were incubated with different lectins before FFE, the one distinct peak representing intact zymogen granules was shifted towards the cathode in the case of concanavalin A (Con A) and Ricinus communis agglutinin 120 (RCA 120). No splitting of the peak occurred. Our results do not support the hypothesis of a coexistence of more than one distinct population of zymogen granules.
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PMID:Separation and analysis of pig pancreatic zymogen granules with free flow electrophoresis and lectins. 792 32

Among the biological exposure indices of lead, lead in plasma was the most direct indicator of current exposure. Lead mobilized into plasma as well as in urine could be used as an indicator of the internal dose of lead. The ratio of non-treated to restored activity of delta-aminolevulinic acid dehydratase (ALA-D) was a more specific index than ALA-D activity itself at low levels of lead exposure, excluding the familial or genetic variation in the activity. The methods using HPLC for determining heme intermediate improved the evaluation of the lead effect: delta-aminolevulinic acid in plasma, blood, and urine (ALA-P, ALA-B, and ALA-U), coproporphyrin in urine, and zinc protoporphyrin in blood (ZP). ROC (Receiver operating characteristic) curve analyses indicated that the diagnostic values for lead exposure decreased in the order ALA-D ratio > ALA-D activity = ALA-P > ALA-U = ZP. Pyrimidine 5'-nucleotidase activity or pyrimidine nucleotide concentrations in blood was also useful for the monitoring or diagnosis of lead intoxication. Using the HPLC method with inclusion compounds in the mobile phase, hippuric acid, methylhippuric acids, mandelic acid and phenylglyoxylic acid could be simultaneously determined in the urine of workers exposed to a mixture of toluene, xylenes, and ethylbenzene. The correction of the urinary metabolite concentration for specific gravity or creatinine allowed the more specific evaluation of the solvent exposure. In the biological monitoring of chlorinated hydrocarbons such as trichloroethylene, prolonged excretion of the metabolites resulted in a bias between metabolite concentrations and TWA levels of the solvent in a day. The background levels of 2,5-hexanedione (HD) were affected by acid hydrolysis conditions, age, sex and lipid metabolism. Substances hydrolyzed to HD in urine from non-exposed subjects were different from HD detected in the workers exposed to n-hexane. Urinary concentrations of N-acetyl-S-(N-methylcarbamoyl) cysteine (AMCC) served as an index of the average exposure to N, N-dimethylformamide during several preceding work days and may indicate the internal dose, while N-methylformamide may be an index of daily exposure. A simple and rapid method for the determination of urinary alkoxyacetic acids was recently developed for the biological monitoring of workers exposed to glycolethers and their acetates. Urinary butoxy acetic acid (free plus conjugated ones) could be simply determined by gaschromatography after acid hydrolysis of urine. The urinary acetone or methanol concentration determined by the head space technique was also useful for the biological monitoring of workers exposed to isopropanol and/or acetone, or methanol, respectively. Evaluation of exposure to the solvents described above could be carried out by comparing the urinary metabolite concentrations with reference values and the biological exposure index values which were defined as the urinary metabolite concentration corresponding to the threshold value for each solvent.
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PMID:[Studies on the evaluation of exposure to industrial chemicals]. 868 99

Alcohol abuse is an acute health problem throughout the world and alcohol consumption is linked to the occurrence of several pathological conditions. Here we tested the acute effects of ethanol on NTPDases (nucleoside triphosphate diphosphohydrolases) and 5'-nucleotidase in zebrafish (Danio rerio) brain membranes. The results have shown a decrease on ATP (36.3 and 18.4%) and ADP (30 and 20%) hydrolysis after 0.5 and 1% (v/v) ethanol exposure during 60 min, respectively. In contrast, no changes on 5'-nucleotidase activity were observed in zebrafish brain membranes. Ethanol in vitro did not alter ATP and ADP hydrolysis, but AMP hydrolysis was inhibited at 0.5, and 1% (23 and 28%, respectively). Acetaldehyde in vitro, in the range 0.5-1%, inhibited ATP (40-85%) and ADP (28-65%) hydrolysis, whereas AMP hydrolysis was reduced (52, 58 and 64%) at 0.25, 0.5 and 1%, respectively. Acetate in vitro did not alter these enzyme activities. Semi-quantitative expression analysis of NTPDase and 5'-nucleotidase were performed. Ethanol treatment reduced NTPDase1 and three isoforms of NTPDase2 mRNA levels. These findings demonstrate that acute ethanol intoxication may influence the enzyme pathway involved in the degradation of ATP to adenosine, which could affect the responses mediated by adenine nucleotides and nucleosides in zebrafish central nervous system.
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PMID:Ethanol and acetaldehyde alter NTPDase and 5'-nucleotidase from zebrafish brain membranes. 1769 55