Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.16.2 (PCP)
 3,761 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

ATP promoted biphasic effects on both basal and fMLP-stimulated arachidonic acid (AA) release in neutrophil-like HL60 cells: stimulation in the micromolar range (EC50 = 3.2 +/- 0.9 microM) and inhibition at higher concentrations (EC50 = 90 +/- 11 microM). ATP also inhibited UTP- and platelet activating factor-stimulated AA release. Only stimulatory effects of ATP on basal or fMLP-stimulated phospholipase C were observed. The inhibitory effect of ATP on AA release was not due to reacylation of released AA, chelation of extracellular Ca2+, cell permeabilization, or changes in the rise of [Ca2+]i induced by agonist. The inhibition was rapid, being detected within 5-15 s. The inhibitory effect of ATP on fMLP-stimulated AA release could be desensitized by pretreatment of the cells with 2 mM ATP, but not 20 microM ATP, the concentration that resulted in maximal release of AA and inositol phosphates. The inhibition by ATP was neither dependent on generation of adenosine by ATP hydrolysis nor the result of direct interaction of ATP with P1 purinergic receptors. Among other nucleotides tested (CTP, GTP, ITP, TTP, XTP, adenosine 5'-(beta,gamma-methylene)triphosphate (AMP-PCP), adenyl-5'-yl imidodiphosphate (AMP-P(NH)P), ADP, adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), and UTP), only UTP and ATP gamma S displayed biphasic effects with potencies and efficacies almost identical to those of ATP. The other nucleotides only exhibited stimulatory effects (EC50 = 60-300 microM). The results are consistent with a model of dual regulation of AA release by two distinct subtypes of P2U receptors in HL60 cells.
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PMID:Dual regulation of arachidonic acid release by P2U purinergic receptors in dibutyryl cyclic AMP-differentiated HL60 cells. 131 16

Human polymorphonuclear neutrophils (PMN) respond to ATP with an elevation in intracellular calcium and a marked enhancement of O2-production in response to stimulation by the chemotactic peptide N'-formyl-Met-Leu-Phe (FMLP). These pertussis toxin-sensitive pathways appear to be mediated by a nucleotide receptor(s) on the surface of human PMN. In the current study, we have examined the binding to intact human PMN of the ATP analog, adenosine 5'-O-(3-thio[35S] triphosphate) [( 35S]ATP gamma S). On the basis of Scatchard analysis, the binding of [35S]ATP gamma S involves at least two sites, one of high and one of low affinity. In the presence of sodium thiophosphate, a compound which did not affect intracellular increases in calcium induced by ATP or N'-formyl-Met-Leu-Phe, a significant fraction of the [35S]ATP gamma S binding was eliminated. This reduction involved both high and low affinity binding of [35S]ATP gamma S and was related to a reduction in numbers of binding sites. The Kd values for the high affinity binding site were unaffected by the presence of sodium thiophosphate, although the low affinity Kd values were numerically increased by 2-fold. In the presence of thiophosphate, [35S]ATP gamma S binding was specific, saturable, and reversible, and was related to a single class of high affinity (Kd = 36 +/- 19 nM) binding sites (184 +/- 144 sites/cell), together with a second class of low affinity (Kd = 1110 +/- 503 nM) binding sites (13,562 +/- 6,851 sites/cells). Competitive binding experiments, based on the ability of nucleotides and ATP analogs to block [35S]ATP gamma S binding to PMN, revealed a rank order of ATP gamma S greater than ATP greater than 2-MeS-ATP = 8-Bromo ATP greater than ADP = ITP greater than AMP-PCP = GTP much greater than CTP. A comparison between the ability of nucleotides to compete with [35S]ATP gamma S binding and their ability to induce a biologic response (elevation of intracellular calcium) revealed a close correlation (r2 = 0.83). These findings support the possibility of a common nucleotide PMN receptor functionally linked to a cellular response which involves increases in intracellular calcium.
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PMID:Adenosine-5'-O-(3-thiotriphosphate) binding to human neutrophils. Evidence for a common nucleotide receptor. 165 77

Micromolar concentrations of copper (Cu2+) and cysteine induce rapid efflux of calcium from sarcoplasmic reticulum (SR) vesicles. This effect appears to be due to a Cu2+-catalyzed oxidation of the added cysteine to a critical sulfhydryl group on the release protein from sarcoplasmic reticulum (J. L. Trimm, G. Salama, and J. J. Abramson (1986) J. Biol. Chem. 261, 16092-16098). The data presented here indicate that adenine nucleotides synergistically stimulate copper/cysteine (oxidation)-induced calcium efflux from SR vesicles. The order of effectiveness in stimulating calcium efflux is ATP greater than AMP-PCP greater than cAMP greater than AMP greater than adenine approximately NAD approximately NADH. Non-adenine-containing nucleotides such as GTP, CTP, UTP, and ITP and the high energy phosphate compound, acetyl phosphate, were ineffective in stimulating oxidation-induced calcium efflux. The relative effectiveness of various adenine nucleotides in stimulating calcium-induced calcium efflux and oxidation-induced calcium efflux are identical, suggesting that a common mode of action is involved when calcium release is triggered by either method. The stimulatory effect of the adenine nucleotides on oxidation-induced efflux is independent of external magnesium concentration and independent of the magnesium gradient across the SR membrane.
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PMID:Adenine nucleotides stimulate oxidation-induced calcium efflux from sarcoplasmic reticulum vesicles. 245 34

1. The whole-cell voltage-clamp technique was used to study the effects of extracellular ATP on smooth muscle cells isolated from the rat vas deferens. 2. ATP (1-200 microM) elicited an inward-rectifying current that was rapid in onset (less than or equal to 100 ms), reached a peak value that depended on [ATP], and desensitized in the continued presence of ATP (half-time approximately 2 s). 3. Cells recovered from desensitization when incubated in the absence of ATP (resensitization half-time approximately 2 min). 4. A comparison was made of the ability of ATP and several of its structural analogues to stimulate inward current at a negative holding potential. ATP was by far the most effective compound among the series ATP, ADP, AMP, adenosine, GTP, UTP and ITP. ADP elicited a current that was 20-25% as large as that produced by ATP, while the other compounds were ineffective at a concentration which produced a maximal ATP response. 5. AMP-CPP (alpha, beta-methylene ATP), AMP-PCP (beta, gamma-methylene ATP), and AMP-PNP (beta, gamma-imido ATP), which are relatively resistant to hydrolysis, were similarly compared to ATP. While none of these analogues elicited a current resembling the ATP-induced current, AMP-CPP and AMP-PNP each produced a small, relatively sustained inward current; AMP-PCP had little or no effect. 6. The ATP-sensitive conductance is cation selective, but does not appear to discriminate strongly between Na+, K+ and Mg2+. 7. Analysis of the fluctuations which accompany the ATP-induced current suggests that ATP controls a population of channels with a unitary current greater than 0.5 pA at -130 mV. 8. The ATP-evoked current discussed in this report may be responsible for the depolarizing effect of ATP previously described in multicellular preparations of the vas deferens.
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PMID:An ATP-sensitive conductance in single smooth muscle cells from the rat vas deferens. 245 75

1. Adult female Culex pipiens and Culiseta inornata have purinergic receptors that respond to extracellular ADP and related compounds. Stimulation of these receptors caused ingestion of artificial diets. Addition of bicarbonate to the saline solvent enhanced the phagostimulatory effect. Saline-bicarbonate was as effective a solvent as blood plasma for Cx. pipiens, and was used in the dose-effect determinations. Ranking of the potencies was: ADP greater than AMP-PNP greater than ATP = AMP greater than AMP-PCP much greater than 2'dAMP greater than 2'dADP greater than 2'dATP. At 1 mM concentration, ITP, GTP, CTP, UTP, c-AMP, 2'AMP, 3'AMP, DPG, or GSH + glucose caused fewer than 50% of the insects to gorge, as did 2'3'dd-ATP, A tetra P, and AMP-CPP at 100 microM. 2. The potency ranking for Cu. inornata was: ADP greater than AMP-PNP greater than ATP greater than AMP-PCP much greater than AMP much greater than AMP-S. The concentrations required to produce the ED50 response (inducing 50% of the test insects to gorge) were much higher than those required for Cx. pipiens; however, saline, not saline-bicarbonate, was used as the solvent. With the exception of the very low potency of AMP for Cu. inornata, the ADP potency index values for the other chemicals tested on both species are similar.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Purinergic reception by culicine mosquitoes. 290 19

Intracellular ATP-dependent Ca2+-sequestration mechanisms were studied in isolated dispersed rat pancreatic acini following treatment with saponin or digitonin to disrupt their plasma membranes. In the presence of 45Ca2+ concentrations less than 10(-6) mol/liter, addition of 5 mmol/liter ATP caused a rapid increase in 45Ca2+ uptake exceeding the control by fivefold. ADP mimicked the ATP effect by 50 to 60%, whereas other nucleotides such as AMP-PNP, AMP-PCP, CTP, UTP, ITP, GTP, cAMP and cGMP did not. Maximal ATP-promoted Ca2+ uptake was obtained at 10(-5) mol/liter Ca2+. Inhibition of Ca2+ uptake by mitochondrial inhibitors was dependent on the Ca2+ concentration, indicating the presence of different Ca2+ storage systems. Whereas the apparent half-saturation constant found for mitochondrial Ca2+ uptake was approximately 4.5 X 10(-7) mol/liter, in the presence of antimycin and oligomycin (nonmitochondrial uptake) it was approximately 1.4 X 10(-8) mol/liter. In the absence of Mg2+ both ATP- and ADP-promoted Ca2+ uptake was nearly abolished. The Ca2+ ionophore and mersalyl blocked Ca2+ uptake, Electron microscopy showed electron-dense precipitates in the rough endoplasmic reticulum of saponin-treated cells in the presence of Ca2+, oxalate and ATP, which were absent in intact cells and in saponin-cells without ATP or pretreated with A23187. The data suggest the presence of mitochondrial and nonmitochondrial ATP-dependent C2+ storage systems in pancreatic acini. The latter is likely to be located in the rough endoplasmic reticulum.
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PMID:Calcium uptake into acini from rat pancreas: evidence for intracellular ATP-dependent calcium sequestration. 680 Dec 63

We have previously demonstrated in permeabilized rat pancreatic acini that the existence of two affinity states of the pancreatic cholecystokinin (CCK) receptor seen in intact cells depends on the presence of ATP. In the present study, we demonstrate that this effect of ATP is mediated by the enzyme nucleoside diphosphate kinase (NDPK). Northern blot hybridization analysis demonstrated NDPK mRNA in pancreas. Furthermore, pancreatic membranes possessed NDPK activity, which transferred high-energy phosphate groups to [8-3H]GDP. This enzyme also utilized UTP and ITP as a source of gamma-phosphate for GTP formation while guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) was formed in the presence of adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S). However, adenylyl (beta, gamma-methylene)-diphosphate (AMP-PCP) did not serve as a substrate for NDPK. Analysis of 125I-Bolton-Hunter-labeled CCK octapeptide ([125I]BH-CCK-8) binding data in the absence of nucleotides was consistent with a single affinity state with dissociation constant (Kd) equal to 80 pM and maximal binding equal to 50.8 fmol/mg. ATP, UTP, ITP, ATP gamma S, and GTP gamma S all induced two CCK binding affinity states, which in the presence of 1 mM ATP were Kd = 74 pM for high-affinity sites and Kd = 4.3 nM for low-affinity sites: AMP-PCP did not induce two affinity states. GDP at 10 microM had no effect on CCK binding but potentiated the effect of ATP. GTP gamma S, in addition to inducing high- and low-affinity states, also elicited a significant concentration-dependent reduction in the total number of measurable CCK receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Nucleoside diphosphate kinase associated with rat pancreatic membranes regulates CCK receptor affinity. 797 49

Activation of rat basophilic leukaemia cells (RBL-2H3) leads to the secretion of allergic and inflammatory mediators. These cells can be permeabilized, yet still retain their ability to secrete in response to antigen. Secretion can also be induced in permeabilized cells by the addition of the ATP analogue, ATP gamma S [adenosine-5'-O-(3-thiotriphosphate)], which is relatively resistant to phosphatase activity. ATP gamma S-induced secretion (35-50% of total amine) is temperature and concentration-dependent. Calcium enhances secretion, but unlike antigen-induced secretion, it does occur in the absence of calcium and without the requirement for inositol phospholipid hydrolysis. Other ATP analogues induced secretion in the rank order AMP-PNP > or = ATP gamma S >>> AMP-PCP > ATP alpha S = ATP [AMP-PNP, adenylyl-imidodiphosphate; AMP-PCP, adenylyl (beta,gamma-methylene)-diphosphonate; ATP alpha S, adenosine-5'-O-(1-thiotriphosphate)]. At equimolar concentrations, ATP inhibits ATP gamma S-induced secretion by 50%, but prolonged incubation in the presence of ATP gamma S surmounts the ATP inhibition. ADP is nearly as effective an inhibitor, but GTP and ITP are ineffective. It is likely that secretion occurs through attachment at an ATP-binding site, effectively blocking the action of a phosphatase, active later in the normal secretory pathway.
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PMID:Calcium-independent secretion by ATP gamma S from a permeabilized rat basophilic leukaemia cell line (RBL-2H3). 808 86

Reticulomyxa transports particulates, like bacterial and algal prey items, bidirectionally along the outside of its pseudopodia. This cell surface transport and intracellular organelle transport can be reactivated in detergent permeabilized cell models [Orokos et al., 1997: Cell Motil. Cytoskeleton]. We have used this unique system to compare cell surface and organelle mechanochemistry in situ in the same reactivated pseudopodia. The ATPase activities of both types of transport were indistinguishable; each displayed identical nucleoside triphosphate specificity, transport ATPase kinetics, and inhibitor sensitivity. Organelle and cell surface transport reactivation required "hydrolyzable" adenosine nucleoside triphosphates; neither reactivated with GTP, CTP, UTP, ITP, AMP-PNP, AMP-PCP, or ATP-gamma-S. However, other ATP analogues, such as 2'-deoxy-ATP and 3'-deoxy-ATP and 2',3'-dideoxy-ATP, supported the reactivation of organelle and cell surface transport at similar, but markedly reduced, velocities. Both transport processes were inhibited similarly by known inhibitors of dynein ATPases such as erythro-9-(3-[2-hydroxynonyl]) adenine (EHNA) or sodium (Na)-orthovanadate. N-ethylmaleimide (NEM) and ultraviolet (UV) irradiation in the presence of Na-orthovanadate and ATP permanently disabled both transport processes. Organelle and surface transport followed identical Michaelis-Menten kinetics with a calculated Km of 118 microM ATP and a maximum translocation velocity (Vmax) of 8.33 microm/sec. These findings strongly suggest that cell surface transport shares the same cytoplasmic dynein motor [Schliwa et al., 1991: J. Cell Biol. 112:1199-1203] that drives organelle transport.
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PMID:Cell surface and organelle transport share the same enzymatic properties in Reticulomyxa. 938 17

We determined the effect of nucleotides and protein kinase A (PKA) on the Ca(2+)-dependent gating of the cloned intermediate conductance, Ca(2+)-dependent K(+) channel, hIK1. In Xenopus oocytes, during two-electrode voltage-clamp, forskolin plus isobutylmethylxanthine induced a Ca(2+)-dependent increase in hIK1 activity. In excised inside-out patches, addition of ATP induced a Ca(2+)-dependent increase in hIK1 activity (NP(o)). In contrast, neither nonhydrolyzable (AMP-PNP, AMP-PCP) nor hydrolyzable ATP analogs (GTP, CTP, UTP, and ITP) activated hIK1. The ATP-dependent activation of hIK1 required Mg(2+) and was reversed by either exogenous alkaline phosphatase or the PKA inhibitor PKI(5-24). The Ca(2+) dependence of hIK1 activation was best fit with a stimulatory constant (K(s)) of 350 nM and a Hill coefficient (n) of 2.3. ATP increased NP(o) at [Ca(2+)] >100 nM while having no effect on K(s) or n. Mutation of the single PKA consensus phosphorylation site at serine 334 to alanine (S334A) had no effect on the PKA-dependent activation during either two-electrode voltage-clamp or in excised inside-out patches. When expressed in HEK293 cells, ATP activated hIK1 in a Mg(2+)-dependent fashion, being reversed by alkaline phosphatase. Neither PKI(5-24) nor CaMKII(281-309) or PKC(19-31) affected the ATP-dependent activation. Northern blot analysis revealed hIK1 expression in the T84 colonic cell line. Endogenous hIK1 was activated by ATP in a Mg(2+)- and PKI(5-24)-dependent fashion and was reversed by alkaline phosphatase, whereas CaMKII(281-309) and PKC(19-31) had no effect on the ATP-dependent activation. The Ca(2+)-dependent activation (K(s) and n) was unaffected by ATP. In conclusion, hIK1 is activated by a membrane delimited PKA when endogenously expressed. Although the oocyte expression system recapitulates this regulation, expression in HEK293 cells does not. The effect of PKA on hIK1 gating is Ca(2+)-dependent and occurs via an increase in NP(o) without an effect on either Ca(2+) affinity or apparent cooperativity.
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PMID:Kinase-dependent regulation of the intermediate conductance, calcium-dependent potassium channel, hIK1. 1061 55


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