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)

Hydrolysis of phosphatidylcholine by phospholipase D (PLD) leads to the generation of the versatile lipid second messenger, phosphatidic acid (PA), which is involved in fundamental cellular processes, including membrane trafficking, actin cytoskeleton remodeling, cell proliferation and cell survival. PLD activity can be dramatically stimulated by a large number of cell surface receptors and is elaborately regulated by intracellular factors, including protein kinase C isoforms, small GTPases of the ARF, Rho and Ras families and, particularly, by the phosphoinositide, phosphatidylinositol 4,5-bisphosphate (PIP(2)). PIP(2) is well known as substrate for the generation of second messengers by phospholipase C, but is now also understood to recruit and/or activate a variety of actin regulatory proteins, ion channels and other signaling proteins, including PLD, by direct interaction. The synthesis of PIP(2) by phosphoinositide 5-kinase (PIP5K) isoforms is tightly regulated by small GTPases and, interestingly, by PA as well, and the concerted formation of PIP(2) and PA has been shown to mediate receptor-regulated cellular events. This review highlights the regulation of PLD by membrane receptors, and describes how the close encounter of PLD and PIP5K isoforms with small GTPases permits the execution of specific cellular functions.
 ...
PMID:Phospholipase D signaling: orchestration by PIP2 and small GTPases. 1724 4

It has been shown that the activation of G(q)-coupled receptors (G(q)PCRs) in cardiac myocytes inhibits the G protein-gated inwardly rectifying K(+) current (I(GIRK)) via receptor-specific depletion of phosphatidylinositol 4,5-bisphosphate (PIP(2)). In this study, we investigated the mechanism of the receptor-mediated regulation of I(GIRK) in acutely isolated hippocampal CA1 neurons by the muscarinic receptor agonist, carbachol (CCh), and the group I metabotropic glutamate receptor (mGluR) agonist, 3,5-dihydroxyphenylglycine (DHPG). I(GIRK) was activated by the GABA(B) receptor agonist, baclofen. When baclofen was repetitively applied at intervals of 2-3 min, the amplitude of the second I(GIRK) was 92.3 +/- 1.7% of the first I(GIRK) in control. Pretreatment of neurons with CCh or DHPG prior to the second application of baclofen caused a reduction in the amplitude of the second I(GIRK) to 54.8 +/- 1.3% and 51.4 +/- 0.6%, respectively. In PLCbeta1 knockout mice, the effect of CCh on I(GIRK) was significantly reduced, whereas the effect of DHPG remained unchanged. The CCh-mediated inhibition of I(GIRK) was almost completely abolished by PKC inhibitors and pipette solutions containing BAPTA. The DHPG-mediated inhibition of I(GIRK) was attenuated by the inhibition of phospholipase A(2) (PLA(2)), or the sequestration of arachidonic acid. We confirmed that DHPG eliminated the inhibition of I(GIRK) by arachidonic acid. These results indicate that muscarinic inhibition of I(GIRK) is mediated by the PLC/PKC signalling pathway, while group I mGluR inhibition of I(GIRK) occurs via the PLA(2)-dependent production of arachidonic acid. These results present a novel receptor-specific mechanism for crosstalk between G(q)PCRs and GABA(B) receptors.
 ...
PMID:Receptor-specific inhibition of GABAB-activated K+ currents by muscarinic and metabotropic glutamate receptors in immature rat hippocampus. 1725 65

The cholinergic innervation of the prefrontal cortex (PFC) plays a pivotal role in regulating executive functions. Muscarinic receptors activated by acetylcholine depolarize pyramidal neurons in the rodent PFC homologue, but the mechanisms mediating this modulation are controversial. To address this question, we studied the responses of layer V rat pre- and infralimbic cortex pyramidal neurons to muscarinic receptor stimulation. Consistent with previous findings, M(1) receptor stimulation produced a strong depolarization, leading to tonic firing. Voltage-clamp analysis revealed that M(1) activation reduced constitutively active inwardly rectifying (Kir2) K(+) channel currents. Blocking protein kinase C activation or depleting intracellular Ca(2+) stores did not affect the modulation. However, reversal of the modulation was prevented by the phosphoinositide kinase inhibitor, wortmanin, suggesting the modulation was mediated by depletions of membrane phosphatidylinositol-4,5-bisphosphate (PIP(2)). Reduction of Kir2 channel currents by M(1) receptor stimulation significantly increased the temporal summation of excitatory synaptic potentials (EPSPs) evoked by repetitive stimulation of layer I. This action was complimented by M(2/4) receptor mediated presynaptic inhibition of the same terminals. As a consequence of this dual modulation, the responses to a single, isolated afferent volley was reduced, but the response to a high-frequency afferent burst was potentiated.
 ...
PMID:M1 muscarinic receptor modulation of Kir2 channels enhances temporal summation of excitatory synaptic potentials in prefrontal cortex pyramidal neurons. 1737 48

Angiotensin II (ANG II) inhibits bTREK-1 (bovine KCNK2) K(+) channels in bovine adrenocortical cells through a Gq-coupled AT(1) receptor by activation of separate Ca(2+)- and ATP hydrolysis-dependent signaling pathways. Whole cell patch-clamp recording from bovine adrenal zona fasciculata (AZF) cells was used to characterize the ATP-dependent signaling mechanism for inhibition of bTREK-1 by ANG II. We discovered that ATP-dependent inhibition of bTREK-1 by ANG II occurred through a novel mechanism that was independent of PLC and its established downstream effectors. The ATP-dependent inhibition of bTREK-1 by ANG II was not reduced by the PLC antagonists edelfosine and U73122, or by the PKC antagonists bisindolylmaleimide I (BIM) or calphostin C. bTREK-1 was partially inhibited ( approximately 25%) by the PKC activator phorbol 12,13 dibutyrate (PDBu) through an ATP-dependent mechanism that was blocked by BIM. Addition of Phosphatidylinositol(4,5) bisphosphate diC8 [DiC(8)PI(4,5)P(2)], a water-soluble derivative of phosphotidyl inositol 4,5 bisphosphate (PIP(2)) to the pipette solution failed to alter inhibition by ANG II. bTREK-1 inhibition by ANG II was also insensitive to antagonists of other protein kinases activated by ANG II in adrenocortical cells but was completely blocked by inorganic polytriphosphate PPPi. DiC(8)PI(4,5)P(2) was a weak activator of bTREK-1 channels, compared with the high-affinity ATP analog N(6)-(2-phenylethyl)adenosine-5'-O-triphosphate (6-PhEt-ATP). These results demonstrate that the modulation of bTREK-1 channels in bovine AZF cells is distinctive with respect to activation by phosphoinositides and nucleotides and inhibition by Gq-coupled receptors. Importantly, ANG II inhibits bTREK-1 channels through a novel pathway that is different from that described for inhibition of native TREK-1 channels in neurons, or cloned channels expressed in cell lines. They also indicate that, under physiological conditions, ANG II inhibits bTREK-1 and depolarizes AZF cells by two, novel, independent pathways that diverge proximal to the activation of PLC.
 ...
PMID:Angiotensin II inhibits native bTREK-1 K+ channels through a PLC-, kinase C-, and PIP2-independent pathway requiring ATP hydrolysis. 1749 31

We have recently demonstrated the involvement of phospholipase D (PLD) in actin polymerization during mammalian sperm capacitation. In the present study, we investigated the involvement of phosphatidylinositol 3- and 4-kinases (PI3K and PI4K) in actin polymerization, as well as the production of PIP(2(4,5)), which is a known cofactor for PLD activation, during bovine sperm capacitation. PIK3R1 (p85 alpha regulatory subunit of PI3K) and PIKCB (PI4K beta) in bovine sperm were detected by Western blotting and immunocytochemistry. Wortmannin (WT) inhibited PI3K and PI4K type III at concentrations of 10 nM and 10 microM, respectively. PI4K activity and PIP(2(4,5)) production were blocked by 10 microM WT but not by 10 nM WT, whereas PI3K activity and PIP(3(3,4,5)) production were blocked by 10 nM WT. Moreover, spermine, which is a known PI4K activator and a component of semen, activated sperm PI4K, resulting in increased cellular PIP(2(4,5)) and F-actin formation. The increases in PIP(2(4,5)) and F-actin intracellular levels during sperm capacitation were mediated by PI4K but not by PI3K activity. Activation of protein kinase A (PKA) by dibutyryl cAMP enhanced PIP(2(4,5)), PIP(3(3,4,5)), and F-actin formation, and these effects were mediated through PI3K. On the other hand, activation of PKC by phorbol myristate acetate enhanced PIP(2(4,5)) and F-actin formation mediated by PI4K activity, while the PI3K activity and intracellular PIP(3(3,4,5)) levels were reduced. These results suggest that two alternative pathways lead to PI4K activation: indirect activation by PKA, which is mediated by PI3K; and activation by PKC, which is independent of PI3K activity. Our results also suggest that spermine, which is present in the ejaculate, regulates PI4K activity during the capacitation process in vivo.
 ...
PMID:Role of PI3-kinase and PI4-kinase in actin polymerization during bovine sperm capacitation. 1749 16

We showed in our previous study that in hippocampal CA1 neurons the stimulation of muscarinic receptors inhibited the GIRK current (I(GIRK)) via a PLC/PKC pathway, whereas group I metabotropic glutamate receptors (mGluR) inhibited I(GIRK) via a PLA(2)/arachidonic acid pathway. In this study, we present evidence that receptor-mediated signalling pathways activated by the two G(q)-coupled receptors (G(q)PCRs) converge on the inhibition of GIRK channel-PIP(2) interaction. I(GIRK) was activated in acutely isolated hippocampal CA1 neurons by repetitive application of baclofen, a GABA(B) receptor agonist, with a 2-3 min interval. When both CCh and DHPG were pretreated before the second I(GIRK) activation, the magnitude of the second I(GIRK) was 52.2 +/- 2.5% of the first I(GIRK), which was not significantly different from the magnitude of inhibition by CCh or DHPG alone. This result shows that the effects of muscarinic receptor and group I mGluR stimulation on I(GIRK) are not additive but occlusive, suggesting that each pathway may converge to a common mechanism that finally regulates I(GIRK). To test the involvement of PIP(2) in this mechanism, the effect of CCh and DHPG on I(GIRK) was tested in cells loaded with exogenous PIP(2). The inhibition of I(GIRK) by CCh or DHPG was almost completely abolished in PIP(2)-loaded cells. We confirmed that the inhibition of I(GIRK) by direct application of phorbol ester or arachidonic acid was also completely reversed in PIP(2)-loaded cells. These results indicate that the decrease in PIP(2)-channel interactions is the final common mechanism responsible for G(q)PCR-induced inhibitions of I(GIRK) mediated by PKC and arachidonic acid.
 ...
PMID:Decrease in PIP(2) channel interactions is the final common mechanism involved in PKC- and arachidonic acid-mediated inhibitions of GABA(B)-activated K+ current. 1758 38

Heteromultimerization of Kir4.1 and Kir5.1 leads to a channel with distinct functional properties. The heteromeric Kir4.1-Kir5.1 channel is expressed in the eye, kidney and brainstem and has CO(2)/pH sensitivity in the physiological range, suggesting a candidate molecule for the regulation of K(+) homeostasis and central CO(2) chemoreception. It is known that K(+) transport in renal epithelium and brainstem CO(2) chemosensitivity are subject to modulation by hormones and neurotransmitters that activate distinct intracellular signaling pathways. If the Kir4.1-Kir5.1 channel is involved in pH-dependent regulation of cellular functions, it may also be regulated by some of the intracellular signaling systems. Therefore, we undertook studies to determine whether PKC modulates the heteromeric Kir4.1-Kir5.1 channel. The channel expressed using a Kir4.1-Kir5.1 tandem dimer construct was inhibited by the PKC activator PMA in a dose-dependent manner. The channel inhibition was produced via reduction of the P(open). The effect of PMA was abolished by specific PKC inhibitors. In contrast, exposure of oocytes to forskolin (a PKA activator) had no significant effect on Kir4.1-Kir5.1 currents. The channel inhibition appeared to be independent of PIP(2) depletion and PKC-dependent internalization. Several consensus sequences of potential PKC phosphorylation sites were identified in the Kir4.1 and Kir5.1 subunits by sequence scan. Although the C-terminal peptides of both Kir4.1 and Kir5.1 were phosphorylated in vitro, site-directed mutagenesis of individual residues failed to reveal the PKC phosphorylation sites suggesting that the channel may have multiple phosphorylation sites. Taken together, these results suggest that the Kir4.1-Kir5.1 but not the homomeric Kir4.1 channel is strongly inhibited by PKC activation.
 ...
PMID:Protein kinase C dependent inhibition of the heteromeric Kir4.1-Kir5.1 channel. 1758 71

Chemotaxis plays an important role in metastasis of cancer cells. In the current study, we investigated the role of PTEN, a tumor suppressor, in chemotaxis of human breast cancer cells. Over-expression of PTEN inhibited EGF-induced chemotaxis, probably due to an overall reduction of PIP(3) levels. Disruption of PTEN by siRNA caused a marked decrease in chemokinesis, cell adhesion, and membrane spreading, resulting in a severe defect in chemotaxis. In PTEN disrupted cells, PDK1, AKT, and PKCzeta exhibited elevated basal activities, which prevented EGF-induced further activation of these molecules. In the absence of EGF, active PDK1 was detected on multiple directions of the plasma membranes of PTEN disrupted cells, which competed against EGF-induced gradient sensing. To confirm the biological relevance of in vitro studies, both PTEN disrupted cells and its parental human breast cancer cells were injected into tail veins of SCID mice. Mice injected with PTEN disrupted cancer cells showed a marked decrease in lung metastasis. Taken together, our data show that PTEN plays a non-redundant role in EGF-induced chemotaxis of human breast cancer cells, and an optimal level of PTEN is required in these responses.
 ...
PMID:Investigate the role of PTEN in chemotaxis of human breast cancer cells. 1776

Changes in the lipid composition of cardiac myocytes have been reported during cardiac hypertrophy, cardiomyopathy, and infarction. Because a recent study indicates a relation between low phosphatidylinositol-bisphosphate (PIP(2)) levels and reduced intercellular coupling, we tested the hypothesis that agonist-induced changes in PIP(2) can result in a reduction of the functional coupling of cardiomyocytes and, consequently, in changes in conduction velocity. Intercellular coupling was measured by Lucifer Yellow dye transfer in cultured neonatal rat cardiomyocytes. Conduction velocity was measured in cardiomyocytes grown on microelectrode arrays. Intercellular coupling was reduced by angiotensin II (43.7 +/- 9.3%, N = 11) and noradrenaline (58.0 +/- 10.7%, N = 11). To test if reduced intercellular coupling after agonist stimulation was caused by PIP(2)-depletion, myocytes were stimulated by angiotensin II (57.3 +/- 5.7%, N = 14) and then allowed to recover in medium with or without wortmannin (an inhibitor of PIP(2) synthesis). Intercellular coupling fully recovered in control medium (102.1 +/- 8.9%, N = 10), whereas no recovery occurred in the presence of wortmannin (69.3 +/- 7.8%, N = 12). Inhibition of PKC, calmodulin, or arachidonic acid production did not affect the response to either angiotensin II or noradrenaline. Furthermore, decreasing or increasing PIP(2) also decreased and increased intercellular coupling, respectively. This supports the role of PIP(2) in the regulation of intercellular coupling. In beating myocytes, conduction velocity was reduced by angiotensin II stimulation, and recovery after wash out was prevented by inhibition of PIP(2) production. Reductions in PIP(2) inhibit intercellular coupling in cardiomyocytes, and stimulation by physiologically relevant agonists reduces intercellular coupling by this mechanism. The reduction in intercellular coupling lowered conduction velocity.
 ...
PMID:Phosphatidylinositol-bisphosphate regulates intercellular coupling in cardiac myocytes. 1853 30

Levetiracetam (LEV) is an effective antiepileptic drug (AED) with distinct mechanism from the conventional AEDs. The major physiological function of ROMK1 channels is to maintain the resting membrane potential (RMP). In this study, we investigated the mechanisms underling LEV on ROMK1 channels. Xenopus oocytes were injected with mRNA to express the wild-type or mutant ROMK1 channels. Giant inside-out patch clamp recordings were performed to study the effect of LEV on these channels. LEV increased the activity of ROMK1 channels in a concentration-dependent manner and enhanced both wild-type and pH(i) gating residue mutant (K80M) channels over a range of pH(i) values. LEV activated the mutated channels at PIP(2)-binding sites (R188Q, R217A and K218A) and PKC-phosphorylation sites channels (S4A, S183A, T191A, T193A, S201A and T234A). However, this drug failed to enhance the channel activity in the presence of PKA inhibitors and did not activate the mutants of PKA-phosphorylation sites on C-terminal (S219A, S313A) and the constructed mutants (S219D and S313D) that mimic the negative charge carried by a phosphate group bound to a serine. Our results demonstrated PKA-mediated phosphorylation is a novel mechanism for LEV activating ROMK1 channels. These findings show that LEV activates ROMK1 channels independently from pH(i) and not via a PIP(2)- or PKC-dependent pathway. The effects of LEV may come from the PKA-induced conformational change but not charge-charge interaction in ROMK1 channels. Enhancing the activity of ROMK1 channels may be an important molecular mechanism for the antiepileptic effects of LEV in restoring neuronal RMP to prevent seizure spreading.
 ...
PMID:PKA-mediated phosphorylation is a novel mechanism for levetiracetam, an antiepileptic drug, activating ROMK1 channels. 1854 45


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>