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)

The effect of angiotensin II on the biosynthesis of phosphatidylcholine in rat heart myoblastic (H9c2) cells was investigated. Cells were incubated with [methyl-3H]choline, and the labelling of phosphatidylcholine at different time intervals was examined. When cells were pretreated with angiotensin II, a significant increase in the labelling of phosphatidylcholine was observed. Analysis of the labelled phosphatidylcholine precursors indicated that the conversion of phosphocholine to CDP-choline was enhanced by angiotensin II treatment. Determination of enzyme activities in the CDP-choline pathway revealed that the activities of choline kinase or CDP-choline: diacylglycerol cholinephosphotransferase were not changed, but the activities of CTP:phosphocholine cytidylyltransferase were stimulated in both the particulate and soluble fractions. The stimulation of the cytidylyltransferase by angiotensin II was not abolished by okadaic acid, indicating that the activation of the enzyme was not mediated via the okadaic-sensitive dephosphorylation mechanism. Alternatively, the stimulation of the cytidylyltransferase activity was completely abolished by protein kinase C inhibitors. Immunoblotting studies revealed that levels of the cytidylyltransferase in the soluble and particulate fractions were not affected by angiotensin II treatment. We conclude that the increase in phosphatidylcholine biosynthesis by angiotensin II was a direct result of the enhancement of the cytidylyltransferase activity. The enhancement of enzyme activity was not mediated via enzyme translocation, but by a mechanism which was intimately associated with the protein kinase C cascade.
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PMID:The enhancement of phosphatidylcholine biosynthesis by angiotensin II in H9c2 cells. 854 36

Phorbol 12-myristate 13-acetate (PMA), a potent activator of protein kinase C, stimulates both the cellular uptake of radiolabeled choline and the activity of CTP:phosphocholine cytidylyltransferase resulting in increased incorporation of choline into phosphatidylcholine (PtdCho). Because of these multiple actions, it is difficult to determine the contribution of de novo synthesis to PMA-stimulated incorporation of radiolabeled choline into PtdCho. To address this issue, in this work Ha-ras-transformed NIH 3T3 fibroblasts, which were found to readily accumulate radiolabeled choline phosphate from the medium, were used. PMA (100 nM) had no stimulatory effect on the uptake of [14C]choline phosphate by transformed cells, but it significantly (2.0- to 2.9-fold) enhanced the incorporation of this labeled precursor into cellular PtdCho during a 5-h incubation period. A well-detectable (approximately 1.45-fold) stimulatory effect on the incorporation of [14C]choline phosphate into PtdCho was obtained with 10 nM PMA, while maximal effects required 100 nM PMA. In transformed cells, PMA also stimulated incorporation of [14C]choline into PtdCho. However, when [14C]choline phosphate and [14C]choline were directly compared, PMA was found to exert slightly, but consistently, greater stimulatory effects on the incorporation of [14C]choline phosphate into PtdCho. The protein kinase C inhibitor GF 109203X inhibited PMA-induced synthesis of PtdCho from both [14C]choline and [14C]choline phosphate. These results directly demonstrate that PMA can stimulate PtdCho synthesis through the protein kinase C system at a step subsequent to the uptake of choline and the action of choline kinase.
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PMID:Direct proof that phorbol ester accelerates the use of choline phosphate for phosphatidylcholine synthesis in intact cells. 891 50

Hexadecylphosphorylcholine (HePC, D-18506, INN: Mitelfosine) belongs to the family of alkylphosphocholines with anticancer activity. Previous reports have related its antitumoral activity to their ability to interfere with phospholipid metabolism. However a clear mechanism of action has not been established yet. We have investigated the effect of HePC on two enzymes recently reported to play a role in cell growth proliferation, phospholipase D (PLD) and choline kinase (ChoK). Our results demonstrate that treatment with HePC induces a rapid stimulation of PLD, that may be achieved by PKC dependent or independent mechanisms, depending on the cell line investigated. Both PLD1 and PLD2 isoenzymes are sensitive to HePC activation. By contrast, no effect was observed by HePC on ChoK, a new target for anticancer drug development. Furthermore, in all cell lines tested, a chronic exposure of the cells to HePC abrogates PLD activation by either phorbol esters or HePC itself with no effect on total cellular PLD levels. This is reflected in a strong inhibition of PLD activity. We suggest that the inhibitory effects on PLD by HePC may be related to its antitumoral action.
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PMID:Modulation of phospholipase D by hexadecylphosphorylcholine: a putative novel mechanism for its antitumoral activity. 1131 48

The yeast Saccharomyces cerevisiae is a model eukaryotic organism for the study of the regulation of phospholipid synthesis. The major phospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine) are synthesized by complementary (CDP-diacylglycerol and Kennedy) pathways. The regulation of these pathways is complex and is controlled by genetic and biochemical mechanisms. Inositol plays a major role in the regulation of phospholipid synthesis. Inositol-mediated regulation involves the expression of genes and the modulation of enzyme activities. Phosphorylation is a major mechanism by which enzymes and transcription factors are regulated, and indeed, key phospholipid biosynthetic enzymes have been identified as targets of phosphorylation. Protein kinase A phosphorylates CTP synthetase, choline kinase, Mg2+-dependent phosphatidate phosphatase, phosphatidylserine synthase, and the transcription factor Opi1p. CTP synthetase and Opi1p are also phosphorylated by protein kinase C. The phosphorylation of these proteins plays a role in regulating their activities and (or) function in phospholipid synthesis.
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PMID:Phospholipid synthesis in yeast: regulation by phosphorylation. 1505 28

The Saccharomyces cerevisiae CKI1-encoded choline kinase catalyzes the committed step in phosphatidylcholine synthesis via the Kennedy pathway. The enzyme is phosphorylated on multiple serine residues, and some of this phosphorylation is mediated by protein kinase A. In this work we examined the hypothesis that choline kinase is also phosphorylated by protein kinase C. Using choline kinase as a substrate, protein kinase C activity was dose- and time-dependent and dependent on the concentrations of choline kinase (K(m) = 27 microg/ml) and ATP (K(m) = 15 microM). This phosphorylation, which occurred on a serine residue, was accompanied by a 1.6-fold stimulation of choline kinase activity. The synthetic peptide SRSSSQRRHS (V5max/K(m) = 17.5 mm(-1) micromol min(-1) mg(-1)) that contains the protein kinase C motif for Ser25 was a substrate for protein kinase C. A Ser25 to Ala (S25A) mutation in choline kinase resulted in a 60% decrease in protein kinase C phosphorylation of the enzyme. Phosphopeptide mapping analysis of the S25A mutant enzyme confirmed that Ser25 was a protein kinase C target site. In vivo the S25A mutation correlated with a decrease (55%) in phosphatidylcholine synthesis via the Kennedy pathway, whereas an S25D phosphorylation site mimic correlated with an increase (44%) in phosphatidylcholine synthesis. Although the S25A (protein kinase C site) mutation did not affect the phosphorylation of choline kinase by protein kinase A, the S30A (protein kinase A site) mutation caused a 46% reduction in enzyme phosphorylation by protein kinase C. A choline kinase synthetic peptide (SQRRHSLTRQ) containing Ser30 was a substrate (V(max)/K(m) = 3.0 mm(-1) micromol min(-1) mg(-1)) for protein kinase C. Comparison of phosphopeptide maps of the wild type and S30A mutant choline kinase enzymes phosphorylated by protein kinase C confirmed that Ser30 was also a target site for protein kinase C.
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PMID:Phosphorylation of the yeast choline kinase by protein kinase C. Identification of Ser25 and Ser30 as major sites of phosphorylation. 1591 56