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)

Calcitonin gene-related peptide (CGRP), a neuropeptide localized in the cardiac autonomic nervous supply, shares 46% similarity in sequence of amino acids with amylin, a peptide synthesized in pancreatic beta-cells. In the present study, the question was addressed whether these peptides could exert hypertrophic effects in cardiomyocytes isolated from the ventricles of adult rats and maintained in short-term, serum-free primary culture. FCS (10% v/v), employed as a positive control, increased the incorporation of l-[14C]phenylalanine into cellular protein, total content of cellular RNA and total mass of cellular protein significantly. CGRP and amylin also increased each of these parameters significantly and in a concentration-dependent manner; maximum responses occurred at 100 pM and 10 nM for CGRP and amylin, respectively. The selective antagonist at CGRP1-receptors, CGRP8-37(100 nM), inhibited significantly the incorporation of l-[14C] phenylalanine into cellular protein in response to CGRP and amylin. The selective inhibitor of protein kinase C (PKC), bisindolylmalemide (BIM) (5 microM), reduced significantly the incorporation of l-[14C] phenylalanine into cellular protein in response to phenylephrine (1 microM), employed as a positive control, but did not inhibit the response to insulin (1 unit/ml), employed as a negative control. BIM (5 microM) reduced significantly the responses to FCS (10% v/v), amylin (10 nM) and CGRP (10 pM), but did not inhibit the response to CGRP (100 pM). The activity of protein kinase C in membranes prepared from intact myocytes pre-treated for 10 min with the phorbol ester, phorbol 12-myristate 13-acetate (PMA) (100 nM), employed as a positive control, and CGRP (10 pM) was significantly greater than in membranes prepared from cardiomyocytes not subjected to agonist stimulation. Phenylephrine (1 microM) increased significantly the specific activity of creatine kinase but not of lactate dehydrogenase in day 1 cultures of freshly isolated cardiomyocytes. Significant induction of creatine kinase, but not lactate dehydrogenase, was also stimulated by CGRP and amylin; the maximum responses occurred at 100 pM and 100 nM CGRP and amylin, respectively. In conclusion, CGRP and amylin exert hypertrophic effects directly on ventricular cardiomyocytes from the hearts of adult rats in vitro. These effects are: (1) due to de novo protein synthesis since total content of cellular RNA and incorporation of l-[14C]phenylalanine into cellular protein were also increased; (2) mediated by a common population of CGRP1-preferring receptors at which amylin binds with lower potency: (3) mediated, at least partly, by the activation of PKC; (4) may be associated with a fetal shift in gene expression, characterized by selective induction of creatine kinase.
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PMID:Hypertrophic effects of calcitonin gene-related peptide (CGRP) and amylin on adult mammalian ventricular cardiomyocytes. 859 94

Rabbit erythrocytes of progressively increasing age were isolated using an avidin-biotin affinity technique and the activity of protein kinases and other enzymes was analyzed in cytosols and membranes from the isolated cells. The activities of cytosolic protein kinase C (PKC), cAMP-dependent kinase (PKA), and casein kinase type I and II (CKI and II) were all found to undergo an age-dependent decrease of twofold to fourfold over the 8-week lifespan of the cells. Membrane-associated tyrosine kinase showed little or no decrease, but membrane-associated CKI showed a dramatic eightfold decrease over the 8-week period. By contrast, various cytosolic enzymes, including lactate dehydrogenase, phosphoglycerate kinase, pyruvate kinase, and acid phosphatase, showed no change in activity over the same time period. Density-separated human erythrocytes showed qualitatively similar decreases in cytosolic protein kinase activities in the densest fractions, which contain the oldest cells. Our results show that aging erythrocytes undergo progressive loss of protein kinases that may adversely affect various cellular processes. The age-dependent loss of kinase activity reported here is one of the most striking manifestations of erythrocyte senescence yet to be reported.
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PMID:Specific loss of protein kinase activities in senescent erythrocytes. 869 69

A stable hepatoma cell line (L35 cells) showing an activation of the cholesterol 7 alpha-hydroxylase gene (CYP7) that had been silent in the parental hepatoma cell line (H35 cells) was used to examine the influence of bile acids on its gene expression and activity. L35 cells were found to concentrate taurocholate from the culture medium, without any significant effect on the expression of 7 alpha-hydroxylase. At physiologic levels (up to 100 microM), CYP7 mRNA expression was not repressed by any bile acid. At supra-physiologic levels (1 mM), the more hydrophobic dihydroxy bile acids, taurodeoxycholate and taurochenodeoxycholate, decreased CYP7 mRNA without decreasing the relative abundance of beta-actin mRNA. Similar results were obtained by culturing cells with sodium dodecylsulfate (50 microM). The medium of L35 cells treated with either taurochenodeoxycholate (1 mM), taurodeoxycholate (1 mM), or sodium dodecylsulfate (50 microM) contained significantly greater activities of two cytosolic enzymes, lactate dehydrogenase and phosphoglucose isomerase, indicating a cytotoxic response. Activation of protein kinase C by phorbol esters decreased the expression of 7 alpha-hydroxylase mRNA without evidence of cytotoxicity; therefore, the inability of L35 cells to show bile acid repression cannot be ascribed to a lack of an effect by this secondary messenger system. In addition, insulin decreased and dexamethasone increased 7 alpha-hydroxylase mRNA without increasing the release of the cytoplasmic enzyme markers. The combined data suggest that L35 cells are resistant to repression of CYP7 gene expression by bile acids, but display physiologic expression to hormones and protein kinase C activation.
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PMID:Rat hepatoma L35 cells, a liver-differentiated cell line, display resistance to bile acid repression of cholesterol 7 alpha-hydroxylase. 872 21

We have studied the hypothesis that 6,7-dihydroxy-1-methyl-1,2,3,4-tetrahydroisoquinoline (salsolinol) is neurotoxic. Salsolinol induced a significant time and dose related inhibition of 3[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; thiazoyl blue (MTT) reduction, and increased lactate dehydrogenase release (LDH) release from human SH-SY5Y neuroblastoma cells, at concentrations within the range of 1-methyl-4-phenylpyridinium (MPP+) cytotoxicity, in vitro. Cytotoxicity was not inhibited by the addition of antioxidants, monoamine oxidase inhibitors or imipramine. In confluent monolayers, salsolinol stimulated catecholamine uptake with EC50 values of 17 muM and 11 muM, for noradrenaline and dopamine, respectively. Conversely, at concentrations above 100 muM, salsolinol inhibited the uptake of noradrenaline and dopamine, with IC50 values of 411 muM and 379 muM, respectively. The inhibition of catecholamine uptake corresponded to the increase displacement of [3H]nisoxetine from the uptake 1 site by salsolinol, as the Ki (353 muM) for displacement was similar to the IC50 (411 and 379 muM) for uptake. Salsolinol stimulated catecholamine uptake does not involve the uptake recognition site, or elevation of cAMP, cGMP, or inhibition of protein kinase C. Salsolinol also inhibited both carbachol (1 mM) and K+ (100 mM, Na+ adjusted) evoked released of noradrenaline from SH-SY5Y cells, with IC50 values of 500 muM and 120 muM, respectively. In conclusion, salsolinol appears to be cytotoxic to SH-SY5Y cells, via a mechanism that does not require uptake 1, bioactivation by monoamine oxidase, or membrane based free radical damage. The effects of salsolinol on catecholamine uptake, and the mechanism of toxicity require further investigation.
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PMID:Studies on the neurotoxicity of 6,7-dihydroxy-1-methyl-1,2,3,4-tetrahydroisoquinoline (salsolinol) in SH-SY5Y cells. 874 84

We investigated the release of glycosaminoglycans (GAGs) labeled with [3H]glucosamine and [35S]sulfate into the medium from cultured bovine aortic endothelial cells stimulated by phorbol 12-myristate 13-acetate (PMA) which is an activator of protein kinase C (PKC). The GAG release was significantly accelerated by PMA without an increase in the leakage of lactate dehydrogenase but was unchanged by 4 alpha-phorbol 12,13-didecanoate which lacks the ability of PKC activation. The acceleration of GAG release by PMA was strongly suppressed by a PKC inhibitor H-7 but not by HA 1004 which is an inactive analogue of H-7. Characterization of GAGs released into the medium revealed that PMA increased both heparan sulfate and the other GAGs in a similar degree. Although the release of GAGs stimulated by thrombin was also suppressed by another PKC inhibitor staurosporine, stimulation by plasmin was unaffected by the inhibitor. The present data suggest that protein kinase C mediates the release of endothelial cell GAGs including anticoagulant heparan sulfate and the stimulation of the release by thrombin includes this mechanism.
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PMID:Phorbol 12-myristate 13-acetate stimulates the release of glycosaminoglycans from cultured vascular endothelial cells: possible involvement of protein kinase C activation. 877 98

Staurosporine is a potent but non-specific kinase inhibitor. It has served as synthetic template for a variety of analogues, the indolocarbazoles, UCN-01 and CGP 41251, and the bisindolylmaleimides, Ro 31-8220 and GF 109203X, were investigated as growth inhibitors of human-derived A549 human lung adenocarcinoma cells. They were compared with respect to (1) effect on the cell cycle, (2) time dependency of growth arrest and (3) cytotoxic potency. Cells were exposed for 1, 2 and 4 days, or for 6, 12 and 24 h in the case of cycle-synchronised cells, to staurosporine analogues at concentrations at which they inhibited growth by 80% after 4 day exposure. Staurosporine and UCN-01 retarded cells in G0/1, and CGP 41251 appeared to inhibit cell growth without cell cycle specificity. Ro 31-8220 slowed progression of synchronised cells through the cycle; over a longer time period it induced a weak block in G2/M. GF 109203X induced potent G2/M arrest in synchronised cells. This was not so apparent in asynchronous cells, which by day 4 were slowed in G0/1 instead. Growth arrest induced by these inhibitors was more potent after incubation for 4 rather than 2 days. Incubation for 1 day followed by maintenance in drug-free medium for 3 days was sufficient to exert some cytostasis. The differences between cytotoxic and cytostatic concentrations, the former measured by release from cells of lactate dehydrogenase, were 15 000-fold for staurosporine, 300-fold for UCN-01, approximately 400-fold for CGP 41251, 25-fold for Ro 31-8220 and approximately 4-fold for GF 109203X. The results show that PKC-selective staurosporine analogues differ with respect to the mechanisms by which they interfere with the cell cycle. The necessity of long-term exposure for effective growth inhibition and the considerable margin between cytostatic and acute cytotoxic indolocarbazole concentrations are findings which might influence the planning and interpretation of clinical trials of these kinase inhibitors.
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PMID:Differential effects of staurosporine analogues on cell cycle, growth and viability in A549 cells. 888 5

The mechanism for prostaglandin (PG) F2 alpha release from pig endometrium after oxytocin (OT) treatment is unknown. OT may rapidly stimulate inositol (1,4,5)-trisphosphate (IP3) and diacylglycerol (DAG) formation, consistent with the concept of rapid activation of a second-messenger system. In support of this hypothesis, endometrial IP3 levels were increased (P < 0.05) within 0.5 min after treatment with 0.1 microM OT. In contrast, increased DAG formation was not detected after treatment with OT. However, similar to the stimulation of endometrial PGF2 alpha secretion observed after OT treatment (P < 0.001), PGF2 alpha release was increased (P < 0.01) after treatment with phorbol-12-myristate-13-acetate (PMA), which mimics DAG activation of protein kinase C. Further, stimulation of endometrial PGF2 alpha secretion did not result from cell death induced by PMA or OT because lactate dehydrogenase, a cytosolic marker of cellular integrity, did not leak into the medium after PMA or OT treatment. In contrast, 0.5% saponin (positive control for cell death and concomitant release of lactate dehydrogenase) increased PGF2 alpha secretion (P < 0.05) and lactate dehydrogenase release (P < 0.001). These results indicate that OT induces endometrial IP3 production in a rapid manner indicative of a second-messenger system. The finding that increased DAG was not also detected after OT treatment may reflect rapid metabolism or compartmentalized production of DAG involved in the second-messenger stimulation of phospholipase C. The high background of DAG used in the biosynthesis of cellular lipids would obscure the rather small spatially localized changes in DAG levels resulting from the activation of phospholipase C. The finding that DAG was present at approximately 10 to 20-fold higher levels than IP3 in resting cells was consistent with this conclusion.
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PMID:The role of phosphoinositide-derived second messengers in oxytocin-stimulated prostaglandin F2 alpha release from endometrium of pigs. 888 94

Adenosine, synthesized by ecto-5'-nucleotidase, is cardioprotective against ischemia and reperfusion injury. We have previously reported that activation of protein kinase C increases ecto-5'-nucleotidase activity of the rat cardiomyocytes, raising the possibility that activation of protein kinase C protects cardiomyocytes from the irreversible cellular injury via activation of ecto-5'-nucleotidase. To test this hypothesis, cardiomyocytes were isolated from adult male Wistar rats and suspended in modified HEPES-Tyrode buffer solution. The cardiomyocytes were incubated with and without exposure to methoxamine (1 x 10(-6) mol/l) or phorbol 12-myristate 13-acetate (PMA. 1 x 10(-8) mol/l). Ecto-5'-nucleotidase activity increased 15 min after the onset of an exposure to either methoxamine or PMA. Adenosine release during hypoxia and reperfusion was augmented in the methoxamine- and PMA-pretreated cardiomyocytes compared with the untreated cardiomyocytes, which was inhibited by alpha, beta-methyleneadenosine 5'-diphosphate (AOPCP), an inhibitor of ecto-5'-nucleotidase. Irreversible cellular injury assessed by the extent of release of lactate dehydrogenase and the trypan blue exclusion test following 60 min of hypoxia and 60 min of reoxygenation was attenuated in the methoxamine- and PMA-pretreated cardiomyocytes compared with the untreated group, which was also blunted by AOPCP and 8-sulfophenyltheophylline, an adenosine receptor antagonist. An adenosine A1 receptor agonist, N6-cyclohexyladenosine, restored the cardioprotection under the treatment with PMA and AOPCP. We conclude that activation of ecto-5'-nucleotidase via protein kinase C contributes to the attenuation of the irreversible injury of the rat cardiomyocytes due to hypoxia and reoxygenation.
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PMID:Activation of ecto-5'-nucleotidase by protein kinase C attenuates irreversible cellular injury due to hypoxia and reoxygenation in rat cardiomyocytes. 889 53

Gadolinium (Gd3+) is known to be the most paramagnetic ion but is also a very toxic cation often used in pharmacology as a putative stretch-activated channel inhibitor. Gadodiamide, a nonionic Gd3+ chelate, is frequently injected i.v. into magnetic resonance imaging (MRI) to enhance contrast. To determine whether this complex is innocuous in humans, a cytotoxicity study was performed on artificially stimulated human neutrophils (HN). HN were incubated with gadodiamide and with free Gd3+ (GdCl3). The purpose of the study was to estimate possible cell damage after incubation, and to validate further trials based on stimulated cellular models. Measurement of lactate dehydrogenase (LDH) release and the Trypan blue exclusion test were used to assess viability. HN were separated from the blood of healthy volunteers and stimulated by phorbol 12-myristate 13-acetate, a pharmacological reactive which induces protein kinase C activation, superoxide generation, and degranulation by leukotcytes. This study demonstrated that an acellular model is necessary to interpret LDH results. In addition, the experimental conditions of the study demonstrated GdCl3 toxicity on HN viability, while gadodiamide was not harmful.
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PMID:Free gadolinium and gadodiamide, a gadolinium chelate used in magnetic resonance imaging: evaluation of their in vitro effects on human neutrophil viability. 890 Feb 15

We tested the hypothesis that elevation of [Ca2+]i during Ca2+ preconditioning (CPC) is a strong activator of protein kinase C (PKC) and confers unique protection against ischemic injury. CPC consisted of three cycles of Ca2+ depletion (1 minute each) and Ca2+ repletion (5 minutes each). Langendorff-perfused rat hearts were subjected to 40 minutes of global ischemia followed by 30 minutes of reperfusion. Significant functional recovery and decreased lactate dehydrogenase release were observed in CPC hearts compared with ischemic control hearts. In addition, ATP contents were significantly higher and cell structure was better preserved in CPC hearts than in ischemic control hearts. Administration of chelerythrine, a specific PKC inhibitor, completely abolished the CPC-induced cardioprotection. In other groups, in which Ca2+ influx during CPC was inhibited with verapamil, amiloride, and low Na+ perfusion, cardioprotection was significantly reduced. The prominent increase in the membrane PKC activity after CPC was in agreement with immunolocalization of PKC-alpha and PKC-delta in the cell membrane of CPC hearts. These results demonstrate that (1) a transient increase in [Ca2+]i is a prominent feature of CPC and is a strong stimulus for the activation of PKC, (2) the elevation of [Ca2+]i likely occurs via an L-type Ca2+ channel and Na(+)-Ca2+ exchanger, and (3) PKC plays a crucial role in the subcellular mechanisms of protection by CPC.
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PMID:Calcium preconditioning elicits strong protection against ischemic injury via protein kinase C signaling pathway. 892 61


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