1,25-Dihydroxyvitamin D3 and TPA activate phospholipase D in Caco-2 cells: role of PKC-α

1999 ◽  
Vol 276 (4) ◽  
pp. G993-G1004 ◽  
Author(s):  
Sharad Khare ◽  
Marc Bissonnette ◽  
Beth Scaglione-Sewell ◽  
Ramesh K. Wali ◽  
Michael D. Sitrin ◽  
...  
Keyword(s):  
Phospholipase D ◽  
Dihydroxyvitamin D3 ◽  
Dihydroxyvitamin D ◽  
1,25 Dihydroxyvitamin D ◽  
Kinase C ◽  
Pkc Isoforms ◽  
Gf 109203X ◽  
Pkc Β ◽  
Stimulation Of

1,25-Dihydroxyvitamin D3[1,25(OH)2D3] and 12- O-tetradecanoylphorbol 13-acetate (TPA) both activated phospholipase D (PLD) in Caco-2 cells. GF-109203x, an inhibitor of protein kinase C (PKC) isoforms, inhibited this activation by both of these agonists. 1,25(OH)2D3activated PKC-α, but not PKC-β1, -βII, -δ, or -ζ, whereas TPA activated PKC-α, -β1, and -δ. Chronic treatment with TPA (1 μM, 24 h) significantly reduced the expression of PKC-α, -βI, and -δ and markedly reduced the ability of 1,25(OH)2D3or TPA to acutely stimulate PLD. Removal of Ca2+ from the medium, as well as preincubation of cells with Gö-6976, an inhibitor of Ca2+-dependent PKC isoforms, significantly reduced the stimulation of PLD by 1,25(OH)2D3or TPA. Treatment with 12-deoxyphorbol-13-phenylacetate-20-acetate, which specifically activates PKC-βI and -βII, however, failed to stimulate PLD. In addition, the activation of PLD by 1,25(OH)2D3or TPA was markedly reduced or accentuated in stably transfected cells with inhibited or amplified PKC-α expression, respectively. Taken together, these observations indicate that PKC-α is intimately involved in the stimulation of PLD in Caco-2 cells by 1,25(OH)2D3or TPA.

Cross talk between serine/threonine and tyrosine kinases regulates ADP-induced thromboxane generation in platelets

2015 ◽  
Vol 114 (09) ◽  
pp. 558-568 ◽  
Author(s):  
Dheeraj Bhavanasi ◽  
Rachit Badolia ◽  
Bhanu Kanth Manne ◽  
Sumalaxmi Janapati ◽  
Carol Dangelmaier ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Functional Role ◽  
Human Platelets ◽  
Src Family Kinases ◽  
Dependent Manner ◽  
Kinase C ◽  
Pkc Isoforms ◽  
First Time ◽  
Stimulation Of

SummaryADP-induced thromboxane generation depends on Src family kinases (SFKs) and is enhanced with pan-protein kinase C (PKC) inhibitors, but it is not clear how these two events are linked. The aim of the current study is to investigate the role of Y311 phosphorylated PKCδ in regulating ADP-induced platelet activation. In the current study, we employed various inhibitors and murine platelets from mice deficient in specific molecules to evaluate the role of PKCδ in ADP-induced platelet responses. We show that, upon stimulation of platelets with 2MeSADP, Y311 on PKCδ is phosphorylated in a P2Y1/Gq and Lyn-dependent manner. By using PKCδ and Lyn knockout murine platelets, we also show that tyrosine phosphorylated PKCδ plays a functional role in mediating 2MeSADP-induced thromboxane generation. 2MeSADP-induced PKCδ Y311 phosphorylation and thromboxane generation were potentiated in human platelets pre-treated with either a pan-PKC inhibitor, GF109203X or a PKC α/β inhibitor and in PKC α or β knockout murine platelets compared to controls. Furthermore, we show that PKC α/β inhibition potentiates the activity of SFK, which further hyper-phosphorylates PKCδ and potentiates thromboxane generation. These results show for the first time that tyrosine phosphorylated PKCδ regulates ADP-induced thromboxane generation independent of its catalytic activity and that classical PKC isoforms α/β regulate the tyrosine phosphorylation on PKCδ and subsequent thromboxane generation through tyrosine kinase, Lyn, in platelets.

Role of protein kinase C in α1-adrenergic regulation of aNa i in guinea pig ventricular myocytes

2000 ◽  
Vol 279 (4) ◽  
pp. H1661-H1668 ◽  
Author(s):  
Su-Hyun Jo ◽  
Chung-Hyun Cho ◽  
Soo Wan Chae ◽  
Chin O. Lee
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Guinea Pig ◽  
Ventricular Myocytes ◽  
Receptor Activation ◽  
Adrenergic Regulation ◽  
Kinase C ◽  
Gf 109203X ◽  
Stimulation Of

We investigated the role of protein kinase C (PKC) in α1-adrenergic regulation of intracellular Na+activity (aNa i) in single guinea pig ventricular myocytes. aNa i and membrane potentials were measured with the Na+-sensitive indicator sodium-binding benzofuran isophthalate and conventional microelectrodes, respectively, at room temperature (24–26°C) while myocytes were stimulated at a rate of 0.25–0.3 Hz. The PKC activator 4β-phorbol 12-myristate 13-acetate (PMA) decreased aNa i in a concentration-dependent manner. PMA (100 nM) produced a maximal decrease in aNa i of 1.5 mM from 6.5 ± 0.4 to 5.0 ± 0.4 mM (means ± SE, n = 12, P< 0.01). The PMA concentration required for a half-maximal decrease in aNa i was 0.46 ± 0.13 nM ( n = 3, P < 0.01). An inactive phorbol, 4α-phorbol 12-myristate 13-acetate, did not decrease aNa i. The decrease caused by PMA could be blocked by the PKC inhibitors staurosporine and bisindolylmaleimide I (GF-109203X). Stimulation of the α1-adrenoceptor with 50 μM phenylephrine decreased aNa i from 6.1 ± 0.3 to 4.6 ± 0.3 mM ( n = 11, P< 0.01). The decrease in aNa i produced by phenylephrine was blocked by pretreatment with staurosporine, GF-109203X, or PMA. The decrease in aNa i produced by PMA was not prevented by pretreatment with tetrodotoxin but was blocked by pretreatment with strophanthidin or high extracellular K+ concentration. The results suggest that α1-adrenergic receptor activation results in a decrease in aNa i via PKC-induced stimulation of the Na+-K+ pump in cardiac myocytes.

The Role of Insulin in the Stimulation of Renal 1,25-Dihydroxyvitamin D Synthesis by Parathyroid Hormone in Rats*

Endocrinology ◽  
1987 ◽  
Vol 121 (5) ◽  
pp. 1721-1726 ◽  
Author(s):  
KYOJI IKEDA ◽  
TOSHIO MATSUMOTO ◽  
KEIKO MORITA ◽  
HIDEYUKI YAMATO ◽  
HIROO TAKAHASHI ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Dihydroxyvitamin D ◽  
1,25 Dihydroxyvitamin D ◽  
Stimulation Of

Protein kinase C dependent induction of pp125FAK in monocytes by colony stimulating factor-GM: evidence for a synergistic effect by the cytokine and 1,25 dihydroxyvitamin D3

1997 ◽  
Vol 152 (2) ◽  
pp. R19-R22 ◽  
Author(s):  
Raed M Kanan ◽  
Hersha Rathod ◽  
Harish K Datta
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Prolonged Exposure ◽  
Dihydroxyvitamin D3 ◽  
Dihydroxyvitamin D ◽  
Kinase C ◽  
Gf 109203X

Abstract The differentiation of monocytes into osteoclasts has been recently achieved in vitro in a suitable milieu containing morphogens that includes 1,25 dihydroxyvitamin D3, colony stimulating factors, interleukins and the presence of cells of osteoblastic lineage. However, the precise role of these factors in the osteoclastic differentiation process has not yet been examined. Since our previous studies have shown that osteoclasts express a much higher level of focal adhesion kinase (pp125FAK) than cells of macrophage/monocytic lineage, the present study was carried out to ascertain which morphogens are involved in increasing the expression of the kinase during the differentiation of monocytes to osteoclasts. We demonstrate that a marked increase in the expression of pp125FAK occurs only after prolonged exposure to hCSF-GM and combination of hCSF-GM and 1,25 (OH)2 D3. The hCSF-GM was found to be a more potent stimulator of pp125FAK induction than 1,25 (OH)2 D3; interestingly, the presence of both hCSF-GM and 1,25 (OH)2 D3 showed co-operative effect. Furthermore, the presence of a protein kinase C inhibitor, bisindolylmaleimide (GF 109203X), blocked hCSF-GM-mediated induction of focal adhesion kinase, implicating an important role for protein kinase C in the induction of pp125FAK.

Lysophosphatidylcholine stimulates phospholipase D in human coronary endothelial cells: role of PKC

1996 ◽  
Vol 271 (4) ◽  
pp. H1706-H1710 ◽  
Author(s):  
D. A. Cox ◽  
M. L. Cohen
Keyword(s):  
Endothelial Cells ◽  
Protein Kinase ◽  
Phospholipase D ◽  
Kinase Inhibitors ◽  
Protein Kinase Inhibitors ◽  
Human Coronary Artery ◽  
Gf 109203X ◽  
Messenger System ◽  
Stimulation Of

Lysophosphatidylcholine (lyso PC) mediates multiple potentially atherogenic effects on endothelial cells, although the cellular mechanism of these effects remains unclear. Phospholipase D (PLD) has been recognized as a novel second-messenger system that may regulate cellular function. The purpose of this study was to determine the effect of lyso PC on PLD activity in human coronary artery endothelial cells (HCAEC) by measuring [3H]phosphatidylethanol production in cells labeled with [3H]myristic acid. After incubation with lyso PC (20 microM) for 40 min, PLD activity was markedly stimulated from five- to sixfold. Stimulation of PLD activity by lyso PC was concentration dependent (half-maximum effective concentration of 7.6 microM) and was not mimicked by phosphatidylcholine (20 microM). Because PLD can be regulated by protein kinases, the effect of several protein kinase inhibitors on lyso PC-stimulated PLD activity was tested. The protein kinase A inhibitor H-89 (300 nM) and the tyrosine kinase inhibitors genistein (30 microM) and tyrphostin A25 (100 microM) had no effect on the stimulation of PLD by lyso PC (20 microM). The protein kinase C (PKC) inhibitor calphostin C (10-300 nM) affected neither lyso PC (20 microM)-nor 4 beta-phorbol 12,13-dibutyrate (PDBu, 300 nM)-stimulated PLD activity, suggesting that this agent may not inhibit PKC in these cells. In contrast, the selective PKC inhibitors GF-109203X (0.3-10 microM) and chelerythrine (1-30 microM) concentration dependently inhibited lyso PC (20 microM)-stimulated PLD activity and blocked PDBu (300 nM)-stimulated PLD activity. Together, these data document that lyso PC stimulated PLD in human endothelial cells, possibly by a PKC-dependent mechanism, and provide evidence that PLD activation in human endothelium is a novel and important mechanism by which lyso PC mediates its cellular and possibly atherogenic effects.

scholarly journals The role of cytosolic Ca2+, protein kinase C, and protein kinase A in hormonal stimulation of phospholipase D in rat hepatocytes.

1994 ◽  
Vol 269 (2) ◽  
pp. 849-859
Author(s):  
L. Gustavsson ◽  
G. Moehren ◽  
M.E. Torres-Marquez ◽  
C. Benistant ◽  
R. Rubin ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Phospholipase D ◽  
Rat Hepatocytes ◽  
Hormonal Stimulation ◽  
Kinase C ◽  
Kinase A ◽  
Stimulation Of
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