Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.5.4.4 (adenosine deaminase)
5,136 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Despite numerous reports of solubilization of adenosine A1 receptors, little progress has been made in isolating or purifying the receptor, owing to the extreme lability of the preparations. The present solubilization strategies recognized the possible role of endogenous adenosine to produce adenosine-receptor-N-protein complexes, which are intrinsically unstable, and instead attempted to use caffeine to solubilize free adenosine receptors, which might be more stable. Endogenous adenosine was removed from membranes by using adenosine deaminase along with GTP to accelerate the release of receptor-bound adenosine. The receptors were then occupied with caffeine and solubilized with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS) in the presence of glycerol. These soluble preparations exhibited the characteristics of free adenosine receptors. They bound the A1-selective antagonist 8-cyclopentyl-1,3-dipropylxanthine (CPDPX) with high affinity to a single class of binding sites, which were insensitive to GTP. The binding activity was extremely stable, with a half-life of about 5 days at 4 degrees C; there was little change in either receptor number or affinity during 3 days at 4 degrees C. This methodology should greatly facilitate the characterization, isolation and purification of the adenosine A1 receptor.
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PMID:Solubilization of stable adenosine A1 receptors from rat brain. 293 Apr 58

Micromolar concentrations of adenosine and its analogs have profound depressant effects on neuronal firing and synaptic transmission in many brain areas. Using the adenosine agonist 2-chloro[3H]adenosine (Cl[3H]Ado), we have identified a distinct class of micromolar-affinity adenosine binding sites in rat forebrain membranes. Specific Cl[3H]Ado binding was reversible and saturable with an apparent KD of 9.1 microM and a Bmax of 61 pmoles/mg protein. The present studies were conducted using washed brain membrane fractions not treated with adenosine deaminase. Specific Cl[3H]Ado binding under these conditions was insensitive to (-)-N6-(R-phenylisopropyl)adenosine ((-)PIA) and treatment with 3 mM N-ethylmaleimide, unlike high-affinity A1 adenosine receptor binding. Treatment of membranes with adenosine deaminase revealed an additional population of binding sites sensitive to (-)PIA. Inhibition of Cl[3H]Ado binding by adenosine analogs exhibited an order of potency ClAdo greater than 5'-N-ethylcarboxamide adenosine (NECA) greater than (-)PIA which differs from that of both A1 and A2 adenosine receptors. The potent A1 and A2 receptor antagonist 8-phenyltheophylline had no significant effect on binding up to 10 microM. Specific binding, however, was inhibited by the adenosine antagonists 8(p-sulfophenyl)theophylline, isobutylmethylxanthine, theophylline, and caffeine. Micromolar Cl[3H]Ado binding was highly selective for adenosine agonists and antagonists. These results suggest that the micromolar-affinity Cl[3H]Ado binding sites may represent a novel central purinergic receptor, distinct from the A1 and A2 adenosine receptors involved in the regulation of adenylate cyclase.
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PMID:A new class of adenosine receptors in brain. Characterization by 2-chloro[3H]adenosine binding. 300 93

Endogenous adenosine acting via A1 adenosine receptors is capable of inhibiting adenylate cyclase activity and neurotransmitter release in the brain. In this report, we describe the synthesis and attributes of a new series of A1 adenosine receptor agonists. One of these, [125I]N6-2-(4-amino-3-iodophenyl)ethyladenosine, can be used as a radioligand and another, [125I]N6-2-(4-azido-3-iodophenyl)ethyladenosine, as a photoaffinity probe. The unlabeled ligand, N6-2-(4-aminophenyl)ethyladenosine, and its iodinated product are full agonists, inhibiting cyclic AMP production in rat cerebral cortex membranes to the same extent as the prototypic A1 agonist N6-R-1-phenyl-2-propyladenosine. These new ligands are not substrates for adenosine deaminase. The new photoaffinity azide described here labels an Mr 38,000 protein that displays all the pharmacological characteristics expected of the A1 adenosine receptor. This is the same molecular-weight protein previously described using a cross-linking radioligand. This new azide compound demonstrates a 15-fold higher efficiency of incorporation, making it the photoaffinity probe of choice for tissues containing low concentrations of A1 adenosine receptors.
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PMID:Characterization of the A1 adenosine receptor-adenylate cyclase system of cerebral cortex using an agonist photoaffinity ligand. 301 53

Although many of the new cardiotonic agents are known to increase cAMP and to inhibit with variable potency a low Km cAMP phosphodiesterase, there is still debate as to the mechanism(s) by which these agents act. In a rat adipocyte membrane model we demonstrate that only approximately 50% of the effect of the new cardiotonic agent sulmazole on cAMP accumulation can be attributed to phosphodiesterase inhibition and that the remaining production of cAMP involves stimulation of adenylate cyclase activity. Two distinct pathways for stimulation of adenylate cyclase are herein reported. Sulmazole, UD-CG 212 CL, enoximone, piroximone, amrinone, and milrinone are all shown to be competitive antagonists of inhibitory A1 adenosine receptors, with EC50 values of 11-909 microM. Elimination of the effects of endogenous adenosine with adenosine deaminase reveals a third distinct mechanism for activation of adenylate cyclase. This mechanism appears to involve Gi, the inhibitory guanine nucleotide-regulatory protein, in that sulmazole attenuates the capacity of GTP to inhibit adenylate cyclase activity, and covalent modification of Gi by pertussis toxin treatment abolishes the capacity of sulmazole to mediate stimulation. Thus, functional blockade of Gi activity is the likely mode of action. Restoration of sulmazole's stimulatory effect on adenylate cyclase activity in pertussis toxin-treated membranes can be accomplished by reconstituting purified preparations of either Gi or mixtures of Gi/Go into treated adipocyte membranes. Of note, this stimulatory effect is completely reversed by inhibitory receptor agonists. Thus, the new cardiotonic agent sulmazole mediates increases in cAMP accumulation by mechanisms other than phosphodiesterase inhibition, including A1 adenosine receptor antagonism and inhibition of Gi function.
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PMID:The new cardiotonic agent sulmazole is an A1 adenosine receptor antagonist and functionally blocks the inhibitory regulator, Gi. 312 27

XAC, a high affinity antagonist of the A1 adenosine receptor, enhances adenylate cyclase activity by 1.3-2 fold with an EC50 of approximately 47 nM in adipocyte membranes pretreated with adenosine deaminase to eliminate adenosine and in the presence of total phosphodiesterase inhibition by 100 microM papaverine. This effect of XAC is observed only at concentrations of GTP sufficient to activate Gi (approximately 5 x 10(-6) M GTP) and is not evident in the absence or presence of lower GTP concentrations. ADP ribosylation of Gi by pertussis toxin treatment also abolishes this stimulatory action of XAC. Furthermore, in the presence of GTP activation of inhibitory prostaglandin E1 receptors diminishes the stimulatory effect of XAC on adenylate cyclase. In addition, XAC interferes with GTP-mediated inhibition of forskolin-stimulated adenylate cyclase activity in a noncompetitive manner. Finally, XAC is only a weak inhibitor of the low Km cyclic AMP phosphodiesterase, producing approximately 40% inhibition of phosphodiesterase activity at a concentration of 100 microM. These data suggest that XAC increases adenylate cyclase activity in absence of endogenous adenosine by inhibiting tonic Gi activity in a reversible manner.
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PMID:A novel site of action of a high affinity A1 adenosine receptor antagonist. 313 23

Decreases in the extracellular Ca2+ concentration (delta Ca) elicited by a 20 Hz/10 s orthodromic stimulus train were measured with combined ion sensitive/recording electrodes in the CA1 area of rat hippocampal slices. Addition of the selective adenosine A1-receptor antagonist, DPCPX, or adenosine deaminase increased evoked delta Ca in the synaptic and pyramidal cell soma layer by more than 100%. This was accompanied by an earlier generation of population spikes during the train. It is concluded that physiological adenosine concentrations of about 1 microM exert a depressive tonus on synaptic transmission and frequency potentiation and that this effect is mediated via A1-receptors.
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PMID:Physiological modulation by adenosine: selective blockade of A1-receptors with DPCPX enhances stimulus train-evoked neuronal Ca influx in rat hippocampal slices. 320 95

The effect of chronic caffeine treatment on lipolysis in rat epididymal adipose tissue was studied. There was a decrease in body weight, epididymal fat pad weight and mean adipocyte diameter in caffeine-treated rats when compared with control rats. No difference in adipocyte triglyceride content or mean adipocyte weight between control and caffeine-treated rats was observed. Lipolysis in adipocytes induced by adenosine deaminase (1 U/ml) decreased by 35% in caffeine-treated rats. This was accompanied by a 2.5-fold increase in the anti-lipolytic potency of 2-chloroadenosine and an increase of adipocyte adenosine A1 receptor number.
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PMID:Potentiation of the anti-lipolytic effect of 2-chloroadenosine after chronic caffeine treatment. 340 45

When tested under conditions reducing the endogenous production of adenosine (presence of adenosine deaminase (ADA) 1.6 IU/ml; and deoxyadenosine triphosphate (d-ATP), and in the presence of both NaCl and GTP, the ADA-resistant analog phenylisopropyladenosine (PIA) inhibited the adenylate cyclase of several brain tissues. These tissues included: (1) 5 brain areas of adult rats (frontal and parietal cortex, cerebellum cortex, hippocampus and striatum)--hypothalamus and mid-brain adenylate cyclases were not inhibited by PIA; (2) astrocytes in primary cultures prepared from cerebral cortex of newborn mice; and (3) neurons in primary cultures prepared from striata of 15-day-old mouse embryos. The specificity profile of the adenosine receptor involved in the inhibition was determined in astrocytes. It was typical of an A1 adenosine receptor (high affinity of PIA; Ka app: 9 +/- 5 X 10(-9) M (n = 4) compared to the affinity of 5'-N-ethylcarboxamide adenosine (NECA); Ka app: 1.3 +/- 0.6 X 10(-7) M (n = 3). There was an excellent correlation between the affinities of several adenosine agonists and antagonists for A1 receptors coupled with an adenylate cyclase in astrocytes and for the receptors labeled with N6-cyclohexyl-[3H]adenosine in brain cortex. In adult rat striatum as well as in astrocytes and striatal neurons in culture the adenylate cyclase was inhibited by low PIA concentrations through A1 receptors and stimulated by higher concentrations through A2 receptors. In contrast, A2 receptors were not detected in adult rat cerebral cortex. In adult rat striatum, A1 and dopamine receptors coupled with an adenylate cyclase seemed to be located on different cell populations. In contrast, in astrocytes A1 and beta-adrenergic receptors coupled with adenylate cyclase were apparently located on the same cells.
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PMID:Inhibition of brain adenylate cyclase by A1 adenosine receptors: pharmacological characteristics and locations. 630 55

Previous reports on a series of benzoylthiophenes, including PD 81,723 [2-amino-4,5-dimethyl-3-(3-trifluoromethyl-benzoyl) thiophene], have shown specific enhancement of agonist binding at the adenosine A1 receptor. We have studied the effects of two substituted benzoylthiophenes, PD 78,416 (thieno[2,3-c]pyridine-6(5H)-carboxylic acid, 2-amino-3-benzoyl-4,7-dihydro-ethyl ester) and RS-74513-000 [2-amino-4-ethyl-5-methyl-3-(3-trifluoro-methyl-benzoyl) thiophene] on response elicited by adenosine A1 receptors in isolated guinea pig left atrium and ileum. In the electrically paced left atrium, PD 78,416 antagonized negative inotropic effect elicited by the agonist CPA [N6-cyclopentyladenosine] with a pKB value of 6.2 +/- 0.2 (n = 4). At a low concentration which had no antagonistic effect (0.1 microM), PD 78,416 enhanced the effect of CPA. The concentration-response curve to CPA was shifted leftward by 5.1 fold (95% confidence limits 2.4-11.2). In field stimulated isolated ileum, PD 78,416 (0.1, 0.3, 1 microM) did not enhance or antagonize effects of CPA. At concentrations above 1 microM, PD 78,416 decreased electrically induced contraction. This effect was not sensitive to adenosine deaminase and was not antagonized by the A1 antagonist CPX [8-cyclopentyl-1,3-dipropyl-xanthine] (1 microM). Unlike PD 78,416, RS-74513-000 (0.01, 0.1, 1, 3, 10 microM) did not antagonize or enhance effects of CPA in the left atrium. However, effects of CPA in ileum were enhanced by RS-74513-000 (1 and 3 microM).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Enhancement of adenosine A1 receptor functions by benzoylthiophenes in guinea pig tissues in vitro. 747 45

1. Electrical responses to volleys in afferent fibers in the optic tract were recorded in the superficial gray layer of anesthetized rat superior colliculus. A prominent negative wave with 4- to 6-ms peak latency in the upper part of the superficial gray layer and a sharp negative wave with 1.5- to 2-ms peak latency in the lower part of the superficial gray layer were elicited, corresponding to the C2 (upper part of the superficial gray layer) and the C1 (lower part of the superficial gray layer) postsynaptic potentials reported by Sefton. 2. These C1 and C2 waves were depressed by kynurenic acid or quinoxaline dione (DNQX) applied just beside the recording electrode, suggesting that neurotransmission in these pathways is mediated by glutamate. 3. Adenosine (10 microM) injected in the superficial gray layer enhanced both C1 and C2 potentials up to 170 and 140%, respectively. 4. Administration of a potent inhibitor of adenosine deaminase, erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA; 5 mg/kg sc) increased the amplitudes of both C1 and C2 potentials to 125 and 130% of the initial levels, respectively. 5. The extracellular application of adenosine uptake inhibitors, dipyridamole (100 microM) and nitrobenzylthioinosine (NBI; 10 microM) also enhanced postsynaptic potentials. 6. Prior application of L-homocysteine thiolactone (10 microM), a compound that facilitates the incorporation of adenosine into S-adenosylhomocystein and reduces the extracellular concentration of adenosine, attenuated the excitatory action of exogenously applied adenosine. 7. Excitatory effects were also observed upon application of a selective adenosine A1 receptor agonist, N6-cyclohexyladenosine (CHA) or a selective A2 receptor agonist, 2-[4-(2-carboxylethyl)- phenethylamino]-5'N-ethylcarboxamide adenosine hydrochloride (CGS21680). Selective A1 and A2 receptor antagonists, 8-cyclopentyl-1,3-dimethylxanthine (CPT) and 3,7-dimethyl-1-propargylxanthine (DMPX), respectively, failed to suppress the excitatory action by adenosine. However, combined application of these two agents blocked the facilitatory action by adenosine on the excitatory synapses. 8. The application of adenosine (10 microM) to the superficial gray layer via a microdialysis probe increased the glutamate release by approximately 230% of the basal level. Similarly, the administration of EHNA (5 mg/kg sc) enhanced the extracellular glutamate level up to approximately 170%. However, prior application of L-homocysteine thiolactone (10 microM) failed to potentiate the glutamate release by adenosine. 9. This is the first in vivo study to demonstrate an excitatory action of adenosine on synaptic transmission.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Adenosine facilitates in vivo neurotransmission in the superior colliculus of the rat. 750 Jan 64


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