Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.11.13 (protein kinase C)
 49,245 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Macrophages express two distinct types of nucleotide (P2 purinergic) receptors for extracellular ATP: one type induces a Ca(2+)-mobilizing response via the activation of phosphatidylinositol-phospholipase C (PI-PLC) while the second type induces the rapid formation of nonselective pores which are permeated by ions and small (< 1 kDa) organic molecules. We have confirmed the presence of these two ATP receptor types in the BAC1.2F5 murine macrophage cell line and have identified 3'-O-(4-benzoyl)benzoyl-ATP (BzATP) as a selective and potent agonist for the so-called P2z or pore-forming ATP receptor type. Several lines of evidence indicated that occupation of these P2z receptors is also accompanied by a rapid and large increase in the activity of a phosphatidylcholine-selective phospholipase D (PLD) effector enzyme. In cells metabolically labeled with [3H]oleic acid or [3H]glycerol and stimulated in the presence of ethanol, ATP and BzATP induced a severalfold increase in the rate and extent of [3H]phosphatidylethanol (PEt) accumulation. These responses were stimulated only by ATP, BzATP, and ATP gamma S (adenosine 5'-O-(3-thiotriphosphate) with the rank order of potency: BzATP >> ATP > ATP gamma A; there was no response to other adenine nucleotides or to non-adenine nucleotides. Significantly, the ability of P2z receptor agonists to stimulate this PLD activity was not dependent on the presence of extracellular [Ca2+] or elevation of cytosolic [Ca2+]. The inability of ionomycin, gramicidin, digitonin, UTP, platelet-activating factor, or phorbol ester to quantitatively mimic these nucleotide effects suggested that activation of this PLD by P2z receptor agonists was not a secondary response due to: 1) enhanced Ca2+ influx; 2) membrane depolarization; 3) nonselective permeabilization of the plasma membrane; 4) stimulation of Ca(2+)-mobilizing ATP receptors; 5) stimulation of a primary PI-PLC pathway; or 6) activation of protein kinase C. These findings suggest that activation of a novel PLD-based signaling pathway may play an important role in the modulation of macrophage function by pore-forming P2z receptors for extracellular ATP.
 ...
PMID:A novel pathway for the activation of phospholipase D by P2z purinergic receptors in BAC1.2F5 macrophages. 133 Oct 96

Extracellular adenosine triphosphate (ATP), at micromolar/nanomolar concentrations, has been shown to induce significant functional changes in a wide variety of normal and transformed cell types. While ATP can be nonspecifically released from the cytosol of damaged cells, it is also co-packaged in certain exocytotic vesicles/granules containing conventional neurotransmitters and hormones. The diverse biologic responses to ATP appear to be mediated by a variety of so-called P2-purinergic, cell surface receptors that are activated upon binding ATP and other nucleotides. Recent physiologic, biochemical, and pharmacologic studies suggest that there are multiple ATP receptor subtypes. These include: (1) G-protein-coupled ATP receptors, which stimulate inositol phospholipid hydrolysis, Ca2+ mobilization, and activation of protein kinase C; (2) ATP receptors that directly activate nonselective cation channels in the plasma membranes of a variety of excitable cell types; and (3) ATP receptors that, via the rapid induction of surface membranes pores permeable to both ions and endogenous metabolites, can produce cytotoxic or activation responses in T lymphocytes and other immune effector cells. In addition to these functional criteria, these putative ATP receptor subtypes can be distinguished by characteristic selectivities for a variety of structurally modified ATP analogs. Current research is directed towards the identification, isolation, and structural characterization of these receptors by both biochemical and molecular biologic approaches.
 ...
PMID:Signal transduction by P2-purinergic receptors for extracellular ATP. 170 33

In cultured intact LLC-PK1 renal epithelial cells, a nonhydrolyzable ATP analogue, ATP gamma S, inhibits AVP-stimulated cAMP formation. In LLC-PK1 membranes, several ATP analogues inhibit basal, GTP-, forskolin-, and AVP-stimulated adenylate cyclase activity in a dose-dependent manner. The rank order potency of inhibition by ATP analogues suggests that a P2y type of ATP receptor is involved in this inhibition. The compound ATP gamma S inhibits agonist-stimulated adenylate cyclase activity in solubilized and in isobutylmethylxanthine (IBMX) and quinacrine pretreated membranes, suggesting that ATP gamma S inhibition occurs independent of AVP and A1 adenosine receptors and of phospholipase A2 activity. The ATP gamma S inhibition of AVP-stimulated adenylate cyclase activity is not affected by pertussis toxin but is attenuated by GDP beta S, suggesting a possible role for a pertussis toxin insensitive G protein in the inhibition. Exposure of intact LLC-PK cells to ATP gamma S results in a significant increase in protein kinase C activity. However, neither of two protein kinase C inhibitors (staurosporine and H-7) prevents ATP gamma S inhibition of AVP-stimulated adenylate cyclase activity, suggesting that this inhibition occurs by a protein kinase C independent mechanism. These findings suggest the presence of functional P2y purinoceptors coupled to two signal transduction pathways in cultured renal epithelial cells. The effect of P2y purinoceptors to inhibit AVP-stimulated adenylate cyclase activity may be mediated, at least in part, by a pertussis toxin insensitive G protein.
 ...
PMID:ATP receptor regulation of adenylate cyclase and protein kinase C activity in cultured renal LLC-PK1 cells. 185 Jul 60

Extracellular ATP and UTP caused a rapid formation of InsP3, with similar kinetics and dose-dependences. ITP also displayed strong agonistic properties in terms of InsP3 production, whereas CTP was almost inactive. Pretreatment of the cells with pertussis toxin attenuated ATP- and UTP-stimulated InsP3 generation to a comparable extent, indicating that both nucleotides couple to phospholipase C by a pertussis-toxin-sensitive G-protein. Short-term (15 min) treatment of the cells with phorbol 12-myristate 13-acetate (PMA) produced a dose-dependent inhibition of ATP- and UTP-induced InsP3 formation. Furthermore, down-regulation of protein kinase C by long-term (24 h) exposure of the cells to PMA resulted in a comparable potentiation of phosphoinositide hydrolysis by both nucleotides. Preincubation of mesangial cells with ATP or UTP caused a pronounced cross-desensitization of subsequent nucleotide-stimulated InsP3 production. ATP and UTP displayed no additivity in terms of InsP3 formation, when used at maximally effective concentrations. In contrast, the peptide hormone angiotensin II interacted in an additive manner with either nucleotide in stimulating phosphoinositide hydrolysis. Reactive Blue 2, a putative P2y-purinoceptor antagonist, caused a rightward shift of both the ATP and UTP dose-response curves. However, since 2-methylthio-ATP was only a partial agonist in stimulating InsP3 formation, the mesangial-cell ATP receptor appears to be different from a classic P2y-receptor. In summary, these results provide no evidence for separate purino- and pyrimidino-ceptors on mesangial cells. In contrast, ATP and UTP may use a common nucleotide receptor for transducing their signals in mesangial cells.
 ...
PMID:Comparison of extracellular ATP and UTP signalling in rat renal mesangial cells. No indications for the involvement of separate purino- and pyrimidino-ceptors. 217 64

The mechanisms of activation of cytoplasmic phospholipase A2 (cPLA2) are complex and incompletely defined. In Chinese hamster ovary (CHO) cells, receptor stimulation of cPLA2 is due to the interaction of pathways involving the alpha subunits of at least two guanine-nucleotide-binding (G) proteins, G alpha i2 and G alpha q. Activation of cPLA2 is inhibited by pertussis toxin and G alpha i2 mutants. In addition, activation of phospholipase C via G alpha q results in increased intracellular calcium ([Ca2+]i) and activation of protein kinase C, both of which interact with and activate cPLA2. The present study was undertaken to analyze the mechanism of interaction of G alpha i2 with the phospholipase-C-stimulated pathway in the activation of cPLA2. We addressed this question using a dominant negative G alpha i2 mutant, [G203T]G alpha i2, in which Gly203 is mutated to Thr. [G203T]G alpha i2 inhibits ATP receptor activation of cPLA2. The effect of [G203T]G alpha i2 was specific to G alpha i2-activated pathways, as shown by its lack of effect on other purinergic receptor stimulated pathways: ATP stimulation of [Ca2+]i or mitogen-activated protein kinase phosphorylation is unaltered by [G203T]G alpha i2. We addressed the possibility that the activation of cPLA2 by Ca2+ and/or protein kinase C is dependent on G alpha i2. Activation of cPLA2 by the Ca2+ ionophore, ionomycin, was inhibited by 61 +/- 9% (n = 5) in [G203T]G alpha i2-expressing cells; however the ionomycin-induced [Ca2+]i rise was unaffected by [G203T]G alpha i2. Thus, [G203T]G alpha i2. specifically inhibits Ca2+ activation of cPLA2. In contrast, activation of cPLA2 via protein kinase C by phorbol 12-myristate 13-acetate was unaffected by [G203T]G alpha i2. Our results demonstrate that Ca2+ but not phorbol ester activation of cPLA2 in CHO cells is G alpha i2-dependent. The possibility is discussed that G alpha i2 is downstream of Ca2+ but upstream of protein kinase C activation of cPLA2.
 ...
PMID:The guanine-nucleotide-binding protein subunit G alpha i2 is involved in calcium activation of phospholipase A2. Effects of the dominant negative G alpha i2 mutant, [G203T]G alpha i2, on activation of phospholipase A2 in Chinese hamster ovary cells. 760 Oct 96

The effect of ATP on cultured striatal neurons was examined by whole cell voltage clamp recordings. ATP produced outwardly rectifying currents that reversed near the expected equilibrium potential for the potassium ion and the currents were blocked by intracellular Cs+. Purinergic receptor agonists such as ADP, AMP adenosine, and 2-methylthio ATP (2-MeSATP) also evoked similar outward currents. The order of their potencies was ATP >> 2-MeSATP > or = ADP > adenosine > AMP, corresponding to a P2 purinergic receptor. ATP-evoked currents were blocked by a specific protein kinase C (PKC) inhibitor, GF109203X. In addition, the intracellular perfusion of a G-protein inactivator, GDP beta S abolished ATP-induced currents, whereas pertussis toxin (PTX) had no effect on the currents. These results suggest that ATP activates a potassium channel in striatal neurons, which is regulated by protein kinase C (PKC) activation through a P2 purinergic receptor linked to PTX-insensitive G protein.
 ...
PMID:ATP-evoked potassium currents in rat striatal neurons are mediated by a P2 purinergic receptor. 764 29

The 85-kDa cytoplasmic phospholipase A2 (cPLA2) is the major hormone and growth factor-regulated enzyme that catalyzes release of arachidonic acid in mammalian cells. Activation of cPLA2 requires elevation of intracellular Ca2+ and the phosphorylation of the cPLA2 enzyme by mitogen-activated protein (MAP) kinase. Down-regulation of protein kinase C by phorbol esters or pertussis toxin catalyzed ADP-ribosylation of Gi proteins inhibits thrombin and ATP receptor-stimulated MAP kinase and arachidonic acid release, indicating that functional protein kinase C and Gi proteins are required for G protein regulation of arachidonic acid release. A mutant G alpha i2 subunit having Gly203 mutated to Thr (alpha i2G203T) inhibited thrombin and ATP receptor stimulation of arachidonic acid release independent of adenylyl cyclase inhibition, Ca2+ mobilization, and MAP kinase activation. Overexpression of the wild-type alpha i2 polypeptide or the inactive mutant alpha i2G204A (Gly204 mutated to Ala) polypeptide had no effect on thrombin or ATP receptor stimulation of arachidonic acid release. The phenotype observed with expression of the mutant alpha i2G203T polypeptide defines a role for Gi2 in the control of cPLA2 activity and subsequent arachidonic acid release in addition to the regulation of intracellular Ca2+ levels and MAP kinase activity.
 ...
PMID:Expression of a mutant Gi2 alpha subunit inhibits ATP and thrombin stimulation of cytoplasmic phospholipase A2-mediated arachidonic acid release independent of Ca2+ and mitogen-activated protein kinase regulation. 829 38

We have recently identified gonadotropes as target cells for ATP action via ATP receptors of the P2U subtype. The present studies have used gonadotrope-derived alpha T3-1 cells to examine the possible signaling mechanisms subserving ATP action in gonadotropes. Addition of ATP produced a biphasic intracellular Ca2+ (Ca2+i) response: a transient spike followed by a small plateau. Removal of extracellular Ca2+ or depolarization with KCl abolished the plateau but had no effect on the spike. The plateau was also blocked by cadmium or nifedipine but not nickel. Pretreatment with GnRH or thapsigargin but not ryanodine inhibited the subsequent Ca2+i response to ATP. Pertussis toxin had no effect on ATP-induced Ca2+i response, whereas the phospholipase C inhibitor U73122 reduced the response. These observations suggest that the Ca2+i response is mediated by a pertussis toxin-insensitive and phospholipase C-coupled G-protein and reflects Ca2+ release from the GnRH- and thapsigargin-sensitive Ca2+ pool followed by Ca2+ influx through high voltage-gated Ca2+ channels. Activation of these ATP receptors had no apparent effects on the cAMP and cGMP signaling systems. Treatment with ATP-gamma S caused the translocation of protein kinase C (PKC) epsilon but not PKC zeta and PKC alpha to the particulate fraction. These data not only characterize the ATP receptor-mediated intracellular signaling in alpha T3-1 cells and render further evidence for a mediator role for nucleotides in gonadotrope function but also provide the first direct demonstration of PKC translocation by ATP receptors.
 ...
PMID:Effects of extracellular nucleotides in the pituitary: adenosine triphosphate receptor-mediated intracellular responses in gonadotrope-derived alpha T3-1 cells. 853 20

There is strong evidence that ATP acts as an excitatory neurotransmitter in the periphery, yet little is known about fast central ATP-mediated transmission. We report here the molecular cloning of a novel neuronal ionotropic ATP receptor of the P2x subtype (P2x3) isolated from rat brain. This central P2x channel subunit has significant amino acid homology with two recently cloned ATP-gated channels from rat smooth muscle (47%) and pheochromocytoma PC12 cells (37%). P2x3 receptor contains the characteristic 10 conserved cysteines of ATP-gated channels, a putative extracellular region homologous to the Walker type A motif found in various nucleotide-binding proteins, and two potential sites for phosphorylation by protein kinase C. Homomeric receptor P2x3 channels expressed in Xenopus oocytes produce rapid cation-selective purinergic currents that are potentiated by zinc ions and reversibly blocked by the P2x antagonists suramin, Reactive Blue 2, and pyridoxalphosphate-6-axophenyl-2U,4U-disulfonic acid. P2x3-receptor subunit mRNA is found in the Purkinje cells and the granule cells of the cerebellum as well as in CA3 pyramidal cells of the hippocampus that are innervated by zinc-rich axon terminals of mossy fibers. Our results suggest that fast excitatory synaptic transmission mediated by zinc-sensitive ATP-gated channels is widespread in mammalian brain.
 ...
PMID:A novel neuronal P2x ATP receptor ion channel with widespread distribution in the brain. 855 29

Rabbit connecting tubule and cortical collecting duct cells were isolated by immunodissection and cultured to confluence on permeable filters and on glass coverslips. Extracellular ATP dose-dependently reduced transcellular Na+ and Ca2+ transport (half-maximal inhibitory concentration, IC50, of 0.5 +/- 0.2 and 3.2 +/- 0.5 microM), with a maximal inhibition of 57 +/- 5 and 43 +/- 4%, respectively. Purinergic receptor agonists inhibited transport with the following rank order of potency: UTP = ATP > ADP; this suggests involvement of P2u purinoceptors. ATP also caused a dose-dependent (50% effective dose, EC50, of 1.5 +/- 0.2 microM) transient increase in intracellular Ca2+ concentration ([Ca2+]i), which decreased to a sustained elevated level. In the absence of extracellular Ca2+, a similar Ca2+ transient occurred, but the sustained response was abolished. Preloading the cells with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) completely prevented the ATP-induced Ca2+ transients, but not the ATP-induced inhibition of Na+ and Ca2+ absorption. Activation of protein kinase C (PKC) by the cell-permeable diacylglycerol analogue, 1,2-dioctanoyl-en-glycerol, mimicked ATP-induced inhibition of Na+ and Ca2+ absorption. The inhibitory effects of ATP were no longer observed after culturing cells in the presence of phorbol ester (12-O-tetradecanoylphorbol-13-acetate) for 5 days, which resulted in downregulation of cellular PKC activity.
 ...
PMID:Inhibition of Na+ and Ca2+ reabsorption by P2u purinoceptors requires PKC but not Ca2+ signaling. 876 22


1 2 3 Next >>