Involvement of Tyrosine Phosphorylation and Protein Kinase C in the Activation of Phospholipase D by H2O2 in Swiss 3T3 Fibroblasts

Abstract

We have investigated the mechanisms involved in H2O2-mediated phospholipase D (PLD) activation in Swiss 3T3 fibroblasts. In the presence of vanadate, H2O2 induced tyrosine phosphorylation of PLD as well as the platelet-derived growth (PDGF) factor receptor, protein kinase Cα (PKCα), and a 62-kDa protein in rat brain PLD1 (rPLD1) immune complexes. PDGF also induced tyrosine phosphorylation of PLD, but this was abolished by catalase, indicating that it was mediated by H2O2 generation. Interestingly, PLD was found to be constitutively associated with the PDGF receptor and PKCα. Stimulation by H2O2 showed a concentration- and time-dependent tyrosine phosphorylation of the proteins in rPLD1 immunoprecipitates and activation of PLD in the cells. Pretreatment of the cells with the protein-tyrosine kinase inhibitors genistein and herbimycin A resulted in a concentration-dependent inhibition of H2O2-induced tyrosine phosphorylation and PLD activation. Activation of PLD by H2O2 was also inhibited dose-dependently by the PKC inhibitors Ro 31-8220 and calphostin C. Down-regulation of PKC by prolonged treatment with 4β-phorbol 12-myristate 13-acetate also abolished H2O2-stimulated PLD activity. H2O2 or vanadate alone did not induce tyrosine phosphorylation of proteins in the rPLD1 immune complex or PLD activation. Reduction of intracellular H2O2levels by pretreatment of the cells with catalase dramatically abrogated tyrosine phosphorylation of proteins in the rPLD1 immune complex and PLD activation, suggesting the potential role of intracellular H2O2 in H2O2-mediated PLD signaling. Taken together, these results suggest that both protein-tyrosine kinase(s) and protein kinase C participate in H2O2-induced PLD activation in Swiss 3T3 cells. We have investigated the mechanisms involved in H2O2-mediated phospholipase D (PLD) activation in Swiss 3T3 fibroblasts. In the presence of vanadate, H2O2 induced tyrosine phosphorylation of PLD as well as the platelet-derived growth (PDGF) factor receptor, protein kinase Cα (PKCα), and a 62-kDa protein in rat brain PLD1 (rPLD1) immune complexes. PDGF also induced tyrosine phosphorylation of PLD, but this was abolished by catalase, indicating that it was mediated by H2O2 generation. Interestingly, PLD was found to be constitutively associated with the PDGF receptor and PKCα. Stimulation by H2O2 showed a concentration- and time-dependent tyrosine phosphorylation of the proteins in rPLD1 immunoprecipitates and activation of PLD in the cells. Pretreatment of the cells with the protein-tyrosine kinase inhibitors genistein and herbimycin A resulted in a concentration-dependent inhibition of H2O2-induced tyrosine phosphorylation and PLD activation. Activation of PLD by H2O2 was also inhibited dose-dependently by the PKC inhibitors Ro 31-8220 and calphostin C. Down-regulation of PKC by prolonged treatment with 4β-phorbol 12-myristate 13-acetate also abolished H2O2-stimulated PLD activity. H2O2 or vanadate alone did not induce tyrosine phosphorylation of proteins in the rPLD1 immune complex or PLD activation. Reduction of intracellular H2O2levels by pretreatment of the cells with catalase dramatically abrogated tyrosine phosphorylation of proteins in the rPLD1 immune complex and PLD activation, suggesting the potential role of intracellular H2O2 in H2O2-mediated PLD signaling. Taken together, these results suggest that both protein-tyrosine kinase(s) and protein kinase C participate in H2O2-induced PLD activation in Swiss 3T3 cells. phospholipase D protein kinase C platelet-derived growth factor rat brain PLD1 phosphatidylbutanol. Cell-surface receptor-mediated hydrolysis of phosphatidylcholine and other membrane phospholipids by phospholipase D (PLD)1 has been recognized as a mechanism of signal transduction in mammalian cells (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar). PLD hydrolyzes phosphatidylcholine to choline and phosphatidic acid. The latter acts as a second messenger and can be further converted to other messenger molecules, namely 1,2-diacylglycerol and lysophosphatidic acid. PLD activation following interaction of agonists with G protein-coupled receptors and receptors with tyrosine kinase activity has been observed in many cells and tissues (3Klein J. Chalifa V. Liscovitch M. Loffelholz K. J. Neurochem. 1995; 65: 1445-1455Crossref PubMed Scopus (111) Google Scholar, 4Liscovitch M. Cantley L.C. Cell. 1995; 81: 659-662Abstract Full Text PDF PubMed Scopus (248) Google Scholar). Although the precise physiological function of PLD in cells is poorly understood, receptor-stimulated PLD activity has been implicated in a broad range of physiological responses (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar, 3Klein J. Chalifa V. Liscovitch M. Loffelholz K. J. Neurochem. 1995; 65: 1445-1455Crossref PubMed Scopus (111) Google Scholar, 4Liscovitch M. Cantley L.C. Cell. 1995; 81: 659-662Abstract Full Text PDF PubMed Scopus (248) Google Scholar, 5Cockcroft S. Biochim. Biophys. Acta. 1992; 1113: 135-160Crossref PubMed Scopus (243) Google Scholar). These include rapid responses (secretion and superoxide generation) as well as long-term responses (proliferation, differentiation, and immune responses). PLD is known to be activated via multiple pathways involving G proteins, protein kinase C (PKC), protein-tyrosine kinases, and Ca2+ (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar).Recently, H2O2 was reported to stimulate PLD activity in endothelial cells (6Natarajan V. Taher M.M. Roehm B. Parinandi N.L. Schmid H.H.O. Kiss Z. Garcia J.G.N. J. Biol. Chem. 1993; 268: 930-937Abstract Full Text PDF PubMed Google Scholar), NIH 3T3 fibroblasts (7Kiss Z. Anderson W.H. Arch. Biochem. Biophys. 1994; 311: 430-436Crossref PubMed Scopus (52) Google Scholar), and PC12 cells S. J. Neurochem. 1997; PubMed Scopus Google Scholar). as and superoxide have been to be in a of cells with growth and agonists of G and it has been that as second J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google Scholar). was also reported that to H2O2 induced tyrosine phosphorylation of proteins that was by with vanadate J. Biol. Chem. Full Text PDF PubMed Google Scholar). The mechanism was to be in to inhibition of protein-tyrosine J. Biol. Chem. Full Text PDF PubMed Google or activation of protein-tyrosine kinase activity M. Biochem. J. PubMed Scopus Google or H2O2 has been reported to tyrosine phosphorylation of the platelet-derived growth factor (PDGF) receptor, the growth factor receptor, and protein to activation of and S. 1995; PubMed Scopus Google Scholar, K. 1995; PubMed Scopus Google Scholar, J. PubMed Scopus Google Scholar, J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, M. S. M. M. Full Text PDF PubMed Scopus Google Scholar). phosphorylation has also been implicated in PLD activation mediated by growth factor Biochem. J. 1992; PubMed Scopus Google Scholar), PDGF Liscovitch PubMed Scopus Google Scholar), S. S. J. Biol. Chem. 1992; Full Text PDF PubMed Google Scholar, J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar), Biochem. J. 1992; PubMed Scopus Google Scholar), and Biochem. Biophys. 1993; PubMed Scopus Google Scholar). inhibitors of protein-tyrosine as of vanadate stimulate PLD activity by G S. S. J. Biol. Chem. 1992; Full Text PDF PubMed Google and Biochem. J. 1993; PubMed Scopus Google it was reported in to by of vanadate, brain PLD1 was and associated with proteins in cells J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google have and a rat brain phospholipase D (rPLD1) J.H. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, J.H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In the have investigated the of as a tyrosine phosphorylation and activation of rPLD1 in Swiss 3T3 fibroblasts. We that H2O2 tyrosine phosphorylation of PLD as well as the PDGF receptor, and protein of in a rPLD1 immune results suggest that protein-tyrosine kinase and PKC participate in H2O2-mediated PLD activation in Swiss 3T3 has been reported that H2O2 PLD activity in endothelial NIH 3T3 and PC12 cells (6Natarajan V. Taher M.M. Roehm B. Parinandi N.L. Schmid H.H.O. Kiss Z. Garcia J.G.N. J. Biol. Chem. 1993; 268: 930-937Abstract Full Text PDF PubMed Google Scholar, Z. Anderson W.H. Arch. Biochem. Biophys. 1994; 311: 430-436Crossref PubMed Scopus (52) Google Scholar, S. J. Neurochem. 1997; PubMed Scopus Google Scholar), but the poorly In the have the of H2O2 and PDGF protein tyrosine phosphorylation and activation of PLD to the involved in the of the in Swiss 3T3 fibroblasts. have that a protein-tyrosine kinase(s) in PLD activation via mechanism V. S. Chem. and PubMed Scopus Google Scholar). that the of vanadate and H2O2 induced tyrosine phosphorylation of PLD and that PLD was constitutively associated with the PDGF receptor and also by These a that of cells to of vanadate tyrosine phosphorylation of the of PLD and of other proteins in the immune complex J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). Interestingly, vanadate or alone to Swiss 3T3 cells tyrosine phosphorylation of proteins in the rPLD1 immune is in with the known of vanadate protein-tyrosine activity in S. Biochem. Biophys. PubMed Scopus Google Scholar). be to the of vanadate in the that in the of intracellular of vanadate is prolonged or the of vanadate J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google the protein tyrosine phosphorylation induced by H2O2 vanadate was by pretreatment with the protein-tyrosine kinase inhibitors genistein and herbimycin The associated PLD activation was also inhibited by these The of protein tyrosine phosphorylation and PLD activation but showed suggesting a role of protein tyrosine phosphorylation in H2O2-induced PLD tyrosine phosphorylation a H2O2 PLD activation A and the tyrosine phosphorylation be involved in H2O2-induced PLD activation, other mechanisms the A with the of these is that the of the inhibitors tyrosine phosphorylation the of this phosphorylation to PLD activation be a reported that H2O2-induced PLD activation was not to PKC activation in PC12 cells S. J. Neurochem. 1997; PubMed Scopus Google Scholar), PKC inhibitors 31-8220 and calphostin and of PKC suggest that H2O2-induced PLD activation is PKC in Swiss 3T3 cells. a in the pathways H2O2-mediated PLD in these cells. tyrosine phosphorylation of PLD by H2O2 is not well with activation of the activation mechanism involving tyrosine phosphorylation of protein The that is involved is this kinase is in to H2O2 D and and of the of PKC inhibition and the tyrosine phosphorylation of PKC induced by H2O2 in cells has been to be associated with in activity M. S. 1997; PubMed Scopus Google Scholar). it be recognized that H2O2 also PLD by a mechanism involving but not tyrosine a is by the of PKC and the results of a tyrosine kinase with a PKC has been that H2O2 protein-tyrosine activity Biochem. Biophys. 1992; Scopus Google and that protein-tyrosine that is activity and to J. Biol. Chem. 1994; Full Text PDF PubMed Google Scholar). it that a protein-tyrosine is a of H2O2 in the of protein-tyrosine be to in tyrosine phosphorylation in of protein-tyrosine 1992; Scopus Google Scholar). In the presence of is to and J. J. Chem. Scholar), cells. to a role in H2O2 protein tyrosine of the vanadate is to the that with proteins Google Scholar). protein-tyrosine in a to to activity L.C. J. Biol. Chem. Full Text PDF PubMed Google Scholar). In protein-tyrosine S. J.H. K. J. J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google Swiss 3T3 PDGF also induced tyrosine phosphorylation of PLD, and a role H2O2 was by the of catalase tyrosine phosphorylation and activity of in and cells has the of H2O2 growth factor and PDGF signal transduction J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google Scholar, 1995; PubMed Scopus Google Scholar). The growth in H2O2 to intracellular protein-tyrosine a in the and the of the growth The of catalase PLD activation by PDGF is not the of the growth factor to PLD via phospholipase C and PKC in Swiss 3T3 is well (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar), and catalase to inhibition of of the PDGF receptor The of catalase the tyrosine phosphorylation of proteins induced by PDGF is in with with growth factor in cells J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google and role H2O2 in this of the was the that rPLD1 immunoprecipitates the PDGF receptor and PKCα. the PDGF receptor and immunoprecipitates PDGF or H2O2 treatment of the cells did not the of rPLD1 with the PDGF receptor or the of rPLD1 with and and and the in the of These suggest that rPLD1 is constitutively associated with the receptor and the protein kinase and that this is not by tyrosine We have also observed that rPLD1 and associated in cells and that the interaction is by treatment of the cells with S. and J. interaction these proteins is by in J.H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, S. K. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). The of rPLD1 with the PDGF receptor is the that it is not by PDGF is to be this PLD has or J.H. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, S. K. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). the has to be is the H2O2 tyrosine phosphorylation of PLD, this the activation of the as by the in the and the of PKC The that tyrosine phosphorylation of or protein is the activation of PLD further as the of the of PLD with the PDGF Cell-surface receptor-mediated hydrolysis of phosphatidylcholine and other membrane phospholipids by phospholipase D (PLD)1 has been recognized as a mechanism of signal transduction in mammalian cells (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar). PLD hydrolyzes phosphatidylcholine to choline and phosphatidic acid. The latter acts as a second messenger and can be further converted to other messenger molecules, namely 1,2-diacylglycerol and lysophosphatidic acid. PLD activation following interaction of agonists with G protein-coupled receptors and receptors with tyrosine kinase activity has been observed in many cells and tissues (3Klein J. Chalifa V. Liscovitch M. Loffelholz K. J. Neurochem. 1995; 65: 1445-1455Crossref PubMed Scopus (111) Google Scholar, 4Liscovitch M. Cantley L.C. Cell. 1995; 81: 659-662Abstract Full Text PDF PubMed Scopus (248) Google Scholar). Although the precise physiological function of PLD in cells is poorly understood, receptor-stimulated PLD activity has been implicated in a broad range of physiological responses (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar, 3Klein J. Chalifa V. Liscovitch M. Loffelholz K. J. Neurochem. 1995; 65: 1445-1455Crossref PubMed Scopus (111) Google Scholar, 4Liscovitch M. Cantley L.C. Cell. 1995; 81: 659-662Abstract Full Text PDF PubMed Scopus (248) Google Scholar, 5Cockcroft S. Biochim. Biophys. Acta. 1992; 1113: 135-160Crossref PubMed Scopus (243) Google Scholar). These include rapid responses (secretion and superoxide generation) as well as long-term responses (proliferation, differentiation, and immune responses). PLD is known to be activated via multiple pathways involving G proteins, protein kinase C (PKC), protein-tyrosine kinases, and Ca2+ (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar). H2O2 was reported to stimulate PLD activity in endothelial cells (6Natarajan V. Taher M.M. Roehm B. Parinandi N.L. Schmid H.H.O. Kiss Z. Garcia J.G.N. J. Biol. Chem. 1993; 268: 930-937Abstract Full Text PDF PubMed Google Scholar), NIH 3T3 fibroblasts (7Kiss Z. Anderson W.H. Arch. Biochem. Biophys. 1994; 311: 430-436Crossref PubMed Scopus (52) Google Scholar), and PC12 cells S. J. Neurochem. 1997; PubMed Scopus Google Scholar). as and superoxide have been to be in a of cells with growth and agonists of G and it has been that as second J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google Scholar). was also reported that to H2O2 induced tyrosine phosphorylation of proteins that was by with vanadate J. Biol. Chem. Full Text PDF PubMed Google Scholar). The mechanism was to be in to inhibition of protein-tyrosine J. Biol. Chem. Full Text PDF PubMed Google or activation of protein-tyrosine kinase activity M. Biochem. J. PubMed Scopus Google or H2O2 has been reported to tyrosine phosphorylation of the platelet-derived growth factor (PDGF) receptor, the growth factor receptor, and protein to activation of and S. 1995; PubMed Scopus Google Scholar, K. 1995; PubMed Scopus Google Scholar, J. PubMed Scopus Google Scholar, J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, M. S. M. M. Full Text PDF PubMed Scopus Google Scholar). phosphorylation has also been implicated in PLD activation mediated by growth factor Biochem. J. 1992; PubMed Scopus Google Scholar), PDGF Liscovitch PubMed Scopus Google Scholar), S. S. J. Biol. Chem. 1992; Full Text PDF PubMed Google Scholar, J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar), Biochem. J. 1992; PubMed Scopus Google Scholar), and Biochem. Biophys. 1993; PubMed Scopus Google Scholar). inhibitors of protein-tyrosine as of vanadate stimulate PLD activity by G S. S. J. Biol. Chem. 1992; Full Text PDF PubMed Google and Biochem. J. 1993; PubMed Scopus Google it was reported in to by of vanadate, brain PLD1 was and associated with proteins in cells J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). We have and a rat brain phospholipase D (rPLD1) J.H. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, J.H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In the have investigated the of as a tyrosine phosphorylation and activation of rPLD1 in Swiss 3T3 fibroblasts. We that H2O2 tyrosine phosphorylation of PLD as well as the PDGF receptor, and protein of in a rPLD1 immune results suggest that protein-tyrosine kinase and PKC participate in H2O2-mediated PLD activation in Swiss 3T3 cells. has been reported that H2O2 PLD activity in endothelial NIH 3T3 and PC12 cells (6Natarajan V. Taher M.M. Roehm B. Parinandi N.L. Schmid H.H.O. Kiss Z. Garcia J.G.N. J. Biol. Chem. 1993; 268: 930-937Abstract Full Text PDF PubMed Google Scholar, Z. Anderson W.H. Arch. Biochem. Biophys. 1994; 311: 430-436Crossref PubMed Scopus (52) Google Scholar, S. J. Neurochem. 1997; PubMed Scopus Google Scholar), but the poorly In the have the of H2O2 and PDGF protein tyrosine phosphorylation and activation of PLD to the involved in the of the in Swiss 3T3 fibroblasts. have that a protein-tyrosine kinase(s) in PLD activation via mechanism V. S. Chem. and PubMed Scopus Google Scholar). that the of vanadate and H2O2 induced tyrosine phosphorylation of PLD and that PLD was constitutively associated with the PDGF receptor and also by These a that of cells to of vanadate tyrosine phosphorylation of the of PLD and of other proteins in the immune complex J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). Interestingly, vanadate or alone to Swiss 3T3 cells tyrosine phosphorylation of proteins in the rPLD1 immune is in with the known of vanadate protein-tyrosine activity in S. Biochem. Biophys. PubMed Scopus Google Scholar). be to the of vanadate in the that in the of intracellular of vanadate is prolonged or the of vanadate J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google the protein tyrosine phosphorylation induced by H2O2 vanadate was by pretreatment with the protein-tyrosine kinase inhibitors genistein and herbimycin The associated PLD activation was also inhibited by these The of protein tyrosine phosphorylation and PLD activation but showed suggesting a role of protein tyrosine phosphorylation in H2O2-induced PLD tyrosine phosphorylation a H2O2 PLD activation A and the tyrosine phosphorylation be involved in H2O2-induced PLD activation, other mechanisms the A with the of these is that the of the inhibitors tyrosine phosphorylation the of this phosphorylation to PLD activation be a reported that H2O2-induced PLD activation was not to PKC activation in PC12 cells S. J. Neurochem. 1997; PubMed Scopus Google Scholar), PKC inhibitors 31-8220 and calphostin and of PKC suggest that H2O2-induced PLD activation is PKC in Swiss 3T3 cells. a in the pathways H2O2-mediated PLD in these cells. tyrosine phosphorylation of PLD by H2O2 is not well with activation of the activation mechanism involving tyrosine phosphorylation of protein The that is involved is this kinase is in to H2O2 D and and of the of PKC inhibition and the tyrosine phosphorylation of PKC induced by H2O2 in cells has been to be associated with in activity M. S. 1997; PubMed Scopus Google Scholar). it be recognized that H2O2 also PLD by a mechanism involving but not tyrosine a is by the of PKC and the results of a tyrosine kinase with a PKC has been that H2O2 protein-tyrosine activity Biochem. Biophys. 1992; Scopus Google and that protein-tyrosine that is activity and to J. Biol. Chem. 1994; Full Text PDF PubMed Google Scholar). it that a protein-tyrosine is a of H2O2 in the of protein-tyrosine be to in tyrosine phosphorylation in of protein-tyrosine 1992; Scopus Google Scholar). In the presence of is to and J. J. Chem. Scholar), cells. to a role in H2O2 protein tyrosine of the vanadate is to the that with proteins Google Scholar). protein-tyrosine in a to to activity L.C. J. Biol. Chem. Full Text PDF PubMed Google Scholar). In protein-tyrosine S. J.H. K. J. J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google Swiss 3T3 PDGF also induced tyrosine phosphorylation of PLD, and a role H2O2 was by the of catalase tyrosine phosphorylation and activity of in and cells has the of H2O2 growth factor and PDGF signal transduction J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google Scholar, 1995; PubMed Scopus Google Scholar). The growth in H2O2 to intracellular protein-tyrosine a in the and the of the growth The of catalase PLD activation by PDGF is not the of the growth factor to PLD via phospholipase C and PKC in Swiss 3T3 is well (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar), and catalase to inhibition of of the PDGF receptor The of catalase the tyrosine phosphorylation of proteins induced by PDGF is in with with growth factor in cells J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google and role H2O2 in this of the was the that rPLD1 immunoprecipitates the PDGF receptor and PKCα. the PDGF receptor and immunoprecipitates PDGF or H2O2 treatment of the cells did not the of rPLD1 with the PDGF receptor or the of rPLD1 with and and and the in the of These suggest that rPLD1 is constitutively associated with the receptor and the protein kinase and that this is not by tyrosine We have also observed that rPLD1 and associated in cells and that the interaction is by treatment of the cells with S. and J. interaction these proteins is by in J.H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, S. K. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). The of rPLD1 with the PDGF receptor is the that it is not by PDGF is to be this PLD has or J.H. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, S. K. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). the has to be is the H2O2 tyrosine phosphorylation of PLD, this the activation of the as by the in the and the of PKC The that tyrosine phosphorylation of or protein is the activation of PLD further as the of the of PLD with the PDGF has been reported that H2O2 PLD activity in endothelial NIH 3T3 and PC12 cells (6Natarajan V. Taher M.M. Roehm B. Parinandi N.L. Schmid H.H.O. Kiss Z. Garcia J.G.N. J. Biol. Chem. 1993; 268: 930-937Abstract Full Text PDF PubMed Google Scholar, Z. Anderson W.H. Arch. Biochem. Biophys. 1994; 311: 430-436Crossref PubMed Scopus (52) Google Scholar, S. J. Neurochem. 1997; PubMed Scopus Google Scholar), but the poorly In the have the of H2O2 and PDGF protein tyrosine phosphorylation and activation of PLD to the involved in the of the in Swiss 3T3 fibroblasts. have that a protein-tyrosine kinase(s) in PLD activation via mechanism V. S. Chem. and PubMed Scopus Google Scholar). that the of vanadate and H2O2 induced tyrosine phosphorylation of PLD and that PLD was constitutively associated with the PDGF receptor and also by These a that of cells to of vanadate tyrosine phosphorylation of the of PLD and of other proteins in the immune complex J. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). Interestingly, vanadate or alone to Swiss 3T3 cells tyrosine phosphorylation of proteins in the rPLD1 immune is in with the known of vanadate protein-tyrosine activity in S. Biochem. Biophys. PubMed Scopus Google Scholar). be to the of vanadate in the that in the of intracellular of vanadate is prolonged or the of vanadate J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google Scholar). the protein tyrosine phosphorylation induced by H2O2 vanadate was by pretreatment with the protein-tyrosine kinase inhibitors genistein and herbimycin The associated PLD activation was also inhibited by these The of protein tyrosine phosphorylation and PLD activation but showed suggesting a role of protein tyrosine phosphorylation in H2O2-induced PLD tyrosine phosphorylation a H2O2 PLD activation A and the tyrosine phosphorylation be involved in H2O2-induced PLD activation, other mechanisms the A with the of these is that the of the inhibitors tyrosine phosphorylation the of this phosphorylation to PLD activation be Although a reported that H2O2-induced PLD activation was not to PKC activation in PC12 cells S. J. Neurochem. 1997; PubMed Scopus Google Scholar), PKC inhibitors 31-8220 and calphostin and of PKC suggest that H2O2-induced PLD activation is PKC in Swiss 3T3 cells. a in the pathways H2O2-mediated PLD in these cells. tyrosine phosphorylation of PLD by H2O2 is not well with activation of the activation mechanism involving tyrosine phosphorylation of protein The that is involved is this kinase is in to H2O2 D and and of the of PKC inhibition and the tyrosine phosphorylation of PKC induced by H2O2 in cells has been to be associated with in activity M. S. 1997; PubMed Scopus Google Scholar). it be recognized that H2O2 also PLD by a mechanism involving but not tyrosine a is by the of PKC and the results of a tyrosine kinase with a PKC has been that H2O2 protein-tyrosine activity Biochem. Biophys. 1992; Scopus Google and that protein-tyrosine that is activity and to J. Biol. Chem. 1994; Full Text PDF PubMed Google Scholar). it that a protein-tyrosine is a of H2O2 in the of protein-tyrosine be to in tyrosine phosphorylation in of protein-tyrosine 1992; Scopus Google Scholar). In the presence of is to and J. J. Chem. Scholar), cells. to a role in H2O2 protein tyrosine of the vanadate is to the that with proteins Google Scholar). protein-tyrosine in a to to activity L.C. J. Biol. Chem. Full Text PDF PubMed Google Scholar). In protein-tyrosine S. J.H. K. J. J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google Scholar). In Swiss 3T3 PDGF also induced tyrosine phosphorylation of PLD, and a role H2O2 was by the of catalase tyrosine phosphorylation and activity of in and cells has the of H2O2 growth factor and PDGF signal transduction J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google Scholar, 1995; PubMed Scopus Google Scholar). The growth in H2O2 to intracellular protein-tyrosine a in the and the of the growth The of catalase PLD activation by PDGF is not the of the growth factor to PLD via phospholipase C and PKC in Swiss 3T3 is well (1Exton J.H. Physiol. Rev. 1997; 77: 303-320Crossref PubMed Scopus (386) Google Scholar, 2Exton J.H. J. Biol. Chem. 1997; 272: 15579-15582Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar), and catalase to inhibition of of the PDGF receptor The of catalase the tyrosine phosphorylation of proteins induced by PDGF is in with with growth factor in cells J. Biol. Chem. 1997; and Full Text Full Text PDF PubMed Scopus Google and role H2O2 in this of the was the that rPLD1 immunoprecipitates the PDGF receptor and PKCα. the PDGF receptor and immunoprecipitates PDGF or H2O2 treatment of the cells did not the of rPLD1 with the PDGF receptor or the of rPLD1 with and and and the in the of These suggest that rPLD1 is constitutively associated with the receptor and the protein kinase and that this is not by tyrosine We have also observed that rPLD1 and associated in cells and that the interaction is by treatment of the cells with S. and J. interaction these proteins is by in J.H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, S. K. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). The of rPLD1 with the PDGF receptor is the that it is not by PDGF is to be this PLD has or J.H. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, S. K. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). the has to be is the H2O2 tyrosine phosphorylation of PLD, this the activation of the as by the in the and the of PKC The that tyrosine phosphorylation of or protein is the activation of PLD further as the of the of PLD with the PDGF We and in the of this