Bradykinin-stimulated cPLA2phosphorylation is protein kinase C dependent in rabbit CCD cells.

We have used an established cell line of rabbit cortical collecting duct (RCCD) epithelial cells representing a mixed population of principal and intercalated cell types to determine which phospholipase A2(PLA2) enzyme therein is responsible for bradykinin (BK)-stimulated arachidonic acid (AA) release and how its activation is regulated. BK-stimulated AA release was reduced 92% by arachidonyl trifluoromethyl ketone, an inhibitor of cytosolic PLA2(cPLA2). Examination of PLA2 activity in vitro demonstrated that BK stimulation resulted in a greater than twofold increase in PLA2 activity and that this activity was dithiothreitol insensitive and was inhibited by an antibody directed against cPLA2. To determine a possible role for protein kinase C (PKC) in the BK-mediated activation of cPLA2, we used the PKC-specific inhibitor Ro31-8220 and examined its effects on AA release, cPLA2 activity, and phosphorylation. Ro31-8220 reduced BK-stimulated AA release and cPLA2 activity by 51 and 58%, respectively. cPLA2 activity stimulated by phorbol ester [phorbol 12-myristate 13-acetate (PMA)] displayed a similar degree of activation and was associated with an increase in serine phosphorylation identical to that caused by BK. The phosphorylation-induced activation of this enzyme was confirmed by the phosphatase-mediated reversal of both BK- and PMA-stimulated cPLA2 activity. In addition, we have also found that PMA stimulation did not cause a synergistic potentiation of BK-stimulated AA release as did calcium ionophore. This occurred despite membrane PKC activity increasing 93% in response to PMA vs. 42% in response to BK. These data, taken together, indicate that cPLA2 is the enzyme responsible for BK-mediated AA release, and, moreover, they indicate that PKC is involved in the onset responses of cPLA2 to BK.

[1]  K. Burns,et al.  Immortalized rabbit cortical collecting duct cells express AT1 angiotensin II receptors. , 1996, The American journal of physiology.

[2]  G. Guillemette,et al.  Calcium‐calmodulin plays a major role in bradykinin‐induced arachidonic acid release by bovine aortic endothelial cells , 1996, Journal of cellular biochemistry.

[3]  K. Malik,et al.  Involvement of mitogen-activated protein kinase and translocation of cytosolic phospholipase A2 to the nuclear envelope in acetylcholine-induced prostacyclin synthesis in rabbit coronary endothelial cells. , 1996, Molecular pharmacology.

[4]  R. Hébert,et al.  Role of PLA2, PLC, and PLD in bradykinin-induced release of arachidonic acid in MDCK cells. , 1996, The American journal of physiology.

[5]  D. Portilla,et al.  Purification of a Novel Calcium-independent Phospholipase A2 from Rabbit Kidney* , 1996, The Journal of Biological Chemistry.

[6]  S. Jaken.  Protein kinase C isozymes and substrates. , 1996, Current opinion in cell biology.

[7]  P. Insel,et al.  Protein kinase C-dependent activation of cytosolic phospholipase A2 and mitogen-activated protein kinase by alpha 1-adrenergic receptors in Madin-Darby canine kidney cells. , 1996, The Journal of clinical investigation.

[8]  William L. Smith,et al.  Calcium-mediated Translocation of Cytosolic Phospholipase A2 to the Nuclear Envelope and Endoplasmic Reticulum (*) , 1995, The Journal of Biological Chemistry.

[9]  J. Brennand,et al.  Activation of phospholipase A2 by the human endothelin receptor in Chinese hamster ovary cells involves Gi protein-mediated calcium influx. , 1995, Biochemical and biophysical research communications.

[10]  S. Watson,et al.  Cytosolic Phospholipase A2 Is Phosphorylated in Collagen- and Thrombin-stimulated Human Platelets Independent of Protein Kinase C and Mitogen-activated Protein Kinase (*) , 1995, The Journal of Biological Chemistry.

[11]  M. Baccarini,et al.  Role of cytosolic phospholipase A2 in arachidonic acid release of rat-liver macrophages: regulation by Ca2+ and phosphorylation. , 1995, The Biochemical journal.

[12]  F. Santini,et al.  Activation of the mitogen-activated protein kinase/cytosolic phospholipase A2 pathway in a rat mast cell line. Indications of different pathways for release of arachidonic acid and secretory granules. , 1995, Journal of immunology.

[13]  R. Sha’afi,et al.  A Mitogen-Activated Protein Kinase-Independent Pathway Involved in the Phosphorylation and Activation of Cytosolic Phospholipase A2 in Human Neutrophils Stimulated with Tumor Necrosis Factor-α , 1995 .

[14]  Y. Nishizuka.  Protein kinase C and lipid signaling for sustained cellular responses , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[15]  M. Jaye,et al.  Activation of Cytosolic Phospholipase A by Basic Fibroblast Growth Factor via a p42 Mitogen-activated Protein Kinase-dependent Phosphorylation Pathway in Endothelial Cells (*) , 1995, The Journal of Biological Chemistry.

[16]  E. Dennis,et al.  Inhibition of Macrophage Ca-independent Phospholipase A by Bromoenol Lactone and Trifluoromethyl Ketones (*) , 1995, The Journal of Biological Chemistry.

[17]  R. Keogh,et al.  The role of protein kinase C in arachidonic acid release and prostaglandin E production from CHO cells transfected with EGF receptors. , 1994, Biochimica et biophysica acta.

[18]  C. Leslie,et al.  Protein kinase C-dependent and -independent pathways of mitogen-activated protein kinase activation in macrophages by stimuli that activate phospholipase A2. , 1994, The Journal of biological chemistry.

[19]  N. Pattabiraman,et al.  Phospholipase A2 enzymes: regulation and physiological role. , 1994, Biochemical pharmacology.

[20]  D. Riendeau,et al.  Arachidonyl trifluoromethyl ketone, a potent inhibitor of 85-kDa phospholipase A2, blocks production of arachidonate and 12-hydroxyeicosatetraenoic acid by calcium ionophore-challenged platelets. , 1994, The Journal of biological chemistry.

[21]  C. Leslie,et al.  Regulation of phospholipase A2 activation by phosphorylation in mouse peritoneal macrophages. , 1993, The Journal of biological chemistry.

[22]  P. Parker,et al.  Isoenzyme specificity of bisindolylmaleimides, selective inhibitors of protein kinase C. , 1993, The Biochemical journal.

[23]  P. Insel,et al.  Inhibition of expression of protein kinase C alpha by antisense cDNA inhibits phorbol ester-mediated arachidonate release. , 1993, The Journal of biological chemistry.

[24]  R. Sundler,et al.  Phosphorylation and activation of the arachidonate-mobilizing phospholipase A2 in macrophages in response to bacteria. , 1993, European journal of biochemistry.

[25]  R. Mayer,et al.  New insights on mammalian phospholipase A2(s); comparison of arachidonoyl‐selective and ‐nonselective enzymes , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[26]  Roger J. Davis,et al.  cPLA2 is phosphorylated and activated by MAP kinase , 1993, Cell.

[27]  G L Johnson,et al.  Phosphorylation and activation of a high molecular weight form of phospholipase A2 by p42 microtubule-associated protein 2 kinase and protein kinase C. , 1993, The Journal of biological chemistry.

[28]  J. Wijkander,et al.  Regulation of arachidonate‐mobilizing phospholipase A2 by phosphorylation via protein kinase C in macrophages , 1992, FEBS letters.

[29]  L. L. Lin,et al.  Cytosolic phospholipase A2 is coupled to hormonally regulated release of arachidonic acid. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[30]  J. D. Clark,et al.  A novel arachidonic acid-selective cytosolic PLA2 contains a Ca2+-dependent translocation domain with homology to PKC and GAP , 1991, Cell.

[31]  S. Hazen,et al.  Suicide inhibition of canine myocardial cytosolic calcium-independent phospholipase A2. Mechanism-based discrimination between calcium-dependent and -independent phospholipases A2. , 1991, The Journal of biological chemistry.

[32]  S. Mumby,et al.  Metabolic radiolabeling techniques for identification of prenylated and fatty acylated proteins , 1990 .

[33]  J. D. Clark,et al.  Purification of a 110-kilodalton cytosolic phospholipase A2 from the human monocytic cell line U937. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[34]  J. Axelrod,et al.  Receptor-mediated activation of phospholipase A2 and arachidonic acid release in signal transduction. , 1990, Biochemical Society transactions.

[35]  R. Anderson,et al.  Bradykinin activates protein kinase C in cultured cortical collecting tubular cells. , 1989, The American journal of physiology.

[36]  P. Insel,et al.  Defining the role of protein kinase C in epinephrine- and bradykinin-stimulated arachidonic acid metabolism in Madin-Darby canine kidney cells. , 1989, Molecular pharmacology.

[37]  E. Nord,et al.  Differential effects of phorbol esters on PGE2 and bradykinin-induced elevation of [Cai2+] in MDCK cells. , 1989, The American journal of physiology.

[38]  P. Insel,et al.  Phorbol ester and neomycin dissociate bradykinin receptor-mediated arachidonic acid release and polyphosphoinositide hydrolysis in Madin-Darby canine kidney cells. Evidence that bradykinin mediates noninterdependent activation of phospholipases A2 and C. , 1988, The Journal of biological chemistry.

[39]  J. A. Shayman,et al.  Islet activating protein inhibits kinin-stimulated inositol phosphate production, calcium mobilization, and prostaglandin E2 synthesis in renal papillary collecting tubule cells independent of cyclic AMP. , 1987, The Journal of biological chemistry.

[40]  H. Taguchi,et al.  New pteridine substrates for dihydropteridine reductase and horseradish peroxidase. , 1986, The Biochemical journal.

[41]  V. Schuster.  Mechanism of bradykinin, ADH, and cAMP interaction in rabbit cortical collecting duct. , 1985, The American journal of physiology.

[42]  V. Dole,et al.  Microdetermination of long-chain fatty acids in plasma and tissues. , 1960, The Journal of biological chemistry.

[43]  R. Smallridge,et al.  Evidence for the involvement of protein kinase C isoforms in alpha-1 adrenergic activation of phospholipase A2 in FRTL-5 thyroid cells. , 1996, Journal of investigative medicine : the official publication of the American Federation for Clinical Research.

[44]  S. Nigam,et al.  Role of intracellular calcium in the regulation of phospholipase A2 in fMet-Leu-Phe-challenged human polymorph neutrophils. , 1995, Agents and actions. Supplements.

[45]  M. Breyer,et al.  Hormonal signaling and regulation of salt and water transport in the collecting duct. , 1994, Annual review of physiology.

[46]  C. Leslie.  Macrophage phospholipase A2 specific for sn-2-arachidonic acid. , 1990, Methods in enzymology.