Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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D609, an Inhibitor of Phosphatidylcholine-specific Phospholipase C, Inhibits Group IV Cytosolic Phospholipase A2

  • Kang, Mi Sun (Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jung, Sung Yun (Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jung, Kwang Mook (Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Kim, Seok Kyun (Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Ahn, Kyong Hoon (Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Kim, Dae Kyong (Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University)
  • Received : 2008.06.03
  • Accepted : 2008.07.21
  • Published : 2008.11.30
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Abstract

As an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), D609 has been widely used to explain the role of PC-PLC in various signal transduction pathways. This study shows that D609 inhibits group IV cytosolic phospholipase $A_2$ ($cPLA_2$), but neither secretory $PLA_2$ nor a $Ca^{2+}$-dependent $PLA_2$. Dixon plot analysis shows a mixed pattern of noncompetitive and uncompetitive inhibition with $K_i=86.25{\mu}M$ for the $cPLA_2$ purified from bovine spleen. D609 also time- and dose-dependently reduces the release of arachidonic acid from a $Ca^{2+}$- ionophore A23187-stimulated MDCK cells. In the AA release experiment, $IC_{50}$ of D609 was ${\sim375}{\mu}M$, suggesting that this reagent may not enter the cells easily. The present study indicates that the inhibitory effects of D609 on various cellular responses may be partially attributable to the inhibition of $cPLA_2$.

Keywords

Acknowledgement

Supported by : Chung-Ang University

References

  1. Amtmann, E. (1996). The antiviral, antitumoural xanthate D609 is a competitive inhibitor of phosphatidylcholine-specific phospholipase C. Drugs. Exp. Clin. Res. 22, 287-294
  2. Andrei, C., Margiocco, P., Poggi, A., Lotti, L.V., Torrisi, M.R., and Rubartelli, A. (2004). Phospholipases C and A2 control lysosome- mediated IL-1 beta secretion: Implications for inflammatory processes. Proc. Natl. Acad. Sci. USA 101, 9745-9750
  3. Bonventre, J.V., (1992). Phospholipase $A_2$ and signal transduction. J. Am. Soc. Nephrol. 3, 128-150
  4. Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  5. Cai, H., Erhardt, P., Szeberenyi, J., Diaz-Meco, M.T., Johansen, T., Moscat, J., and Cooper, G.M. (1992). Hydrolysis of phosphatidylcholine is stimulated by Ras proteins during mitogenic signal transduction. Mol. Cell. Biol. 12, 5329-5335 https://doi.org/10.1128/MCB.12.12.5329
  6. Cai, H., Erhardt, P., Troppmair, J., Diaz-Meco, M.T., Sithanandam, G., Rapp, U.R., Moscat, J., and Cooper, G.M. (1993). Hydrolysis of phosphatidylcholine couples Ras to activation of Raf protein kinase during mitogenic signal transduction. Mol. Cell. Biol. 13, 7645-7651 https://doi.org/10.1128/MCB.13.12.7645
  7. Cheng, J., Weber, J.D., Baldassare, J.J., and Raben, D.M., (1997). Ablation of Go alpha-subunit results in a transformed phenotype and constitutively active phosphatidylcholine-specific phospholipase C. J. Biol. Chem. 272, 17312-17319 https://doi.org/10.1074/jbc.272.28.17312
  8. Cuschieri, J., Billgren, J., and Maier, R.V. (2006). Phosphatidylcholine- specific phospholipase C (PC-PLC) is required for LPSmediated macrophage activation through CD14. J. Leukoc. Biol. 80, 407-414 https://doi.org/10.1189/jlb.1105622
  9. Diaz-Laviada, I., Larrodera, P., Diaz-Meco, M.T., Cornet, M.E., Guddal, P.H., Johansen, T., and Moscat, J. (1990). Evidence for a role of phosphatidylcholine-hydrolysing phospholipase C in the regulation of protein kinase C by ras and src oncogenes. EMBO. J. 9, 3907-3912
  10. Dieter, P., Kolada, A., Kamionka, S., Schadow, A., and Kaszkin, M. (2002). Lipopolysaccharide-induced release of arachidonic acid and prostaglandins in liver macrophages: regulation by Group IV cytosolic phospholipase $A_2$, but not by Group V and Group IIA secretory phospholipase $A_2$. Cell. Signal. 14, 199-204 https://doi.org/10.1016/S0898-6568(01)00243-1
  11. Exton, J.H. (1994). Phosphatidylcholine breakdown and signal transduction. Biochim. Biophys. Acta 1212, 26-42 https://doi.org/10.1016/0005-2760(94)90186-4
  12. Favaron, M., Manev, H., Siman, R., Bertolino, M., Szekely, A.M., DeErausquin, G., Guidotti, A., and Costa, E. (1990). Downregulation of protein kinase C protects cerebellar granule neurons in primary culture from glutamate-induced neuronal death. Proc. Natl. Acad. Sci. USA 87, 1983-1987
  13. Gijon, M.A., and Leslie, C.C. (1999). Regulation of arachidonic acid release and cytosolic phospholipase $A_2$ activation. J. Leukoc. Biol. 65, 330-336 https://doi.org/10.1002/jlb.65.3.330
  14. Johansen, T., Bjorkoy, G., Overvatn, A., Diaz-Meco, M.T., Traavik, T., and Moscat, J. (1994). NIH 3T3 cells stably transfected with the gene encoding phosphatidylcholine hydrolyzing phospholipase C from Bacillus cereus acquire a transformed phenotype. Mol. Cell. Biol. 14, 646-654 https://doi.org/10.1128/MCB.14.1.646
  15. Kennedy, C.R., Proulx, P.R., and Hebert, R.L. (1996). Role of $PLA_2$,PLC, and PLD in bradykinin-induced release of arachidonic acid in MDCK cells. Am. J. Physiol. 271, C1064-C1072 https://doi.org/10.1152/ajpcell.1996.271.4.C1064
  16. Kennedy, C.R., Hebert, R.L., Do, M.T., and Proulx, P.R. (1997). Bradykinin-stimulated arachidonic acid release from MDCK cells is not protein kinase C dependent. Am. J. Physiol. 273, C1605-1612 https://doi.org/10.1152/ajpcell.1997.273.5.C1605
  17. Kim, D.K., and Bonventre, J.V. (1993). Purification of a 100 kDa phospholipase $A_2$ from spleen, lung and kidney: antiserum raised to pig spleen phospholipase $A_2$ recognizes a similar form in bovine lung, kidney and platelets, and immunoprecipitates phospholipase $A_2$ activity. Biochem. J. 294, 261-270 https://doi.org/10.1042/bj2940261
  18. Kim, D.K., Suh, P.G., and Ryu, S.H. (1991). Purification and some properties of a phospholipase $A_2$ from bovine platelets. Biochem. Biophys. Res. Commun. 174, 189-196 https://doi.org/10.1016/0006-291X(91)90504-Z
  19. Kiss, Z., and Tomono, M. (1995). Compound D609 inhibits phorbol ester-stimulated phospholipase D activity and phospholipase Cmediated phosphatidylethanolamine hydrolysis. Biochim. Biophys. Acta 1259, 105-108 https://doi.org/10.1016/0005-2760(95)00148-6
  20. Li, Y., Maher, P., and Schubert, D. (1998). Phosphatidylcholinespecific phospholipase C regulates glutamate-induced nerve cell death. Proc. Natl. Acad. Sci. USA 95, 7748-7753
  21. Lin, L.L., Wartmann, M., Lin, A.Y., Knopf, J.L., Seth, A., and Davis, R.J. (1993). $cPLA_2$ is phosphorylated and activated by MAP kinase. Cell 72, 269-278 https://doi.org/10.1016/0092-8674(93)90666-E
  22. Luberto, C., and Hannun, Y.A. (1998). Sphingomyelin synthase, a potential regulator of intracellular levels of ceramide and diacylglycerol during SV40 transformation. Does sphingomyelin synthase account for the putative phosphatidylcholine-specific phospholipase C? J. Biol. Chem. 273, 14550-14559 https://doi.org/10.1074/jbc.273.23.14550
  23. Machleidt, T., Kramer, B., Adam, D., Neumann, B., Schutze, S., Wiegmann, K., and Kronke, M. (1996). Function of the p55 tumor necrosis factor receptor "death domain" mediated by phosphatidylcholine- specific phospholipase C. J. Exp. Med. 184, 725-733 https://doi.org/10.1084/jem.184.2.725
  24. Macrez-Lepretre, N., Morel, J.L., and Mironneau, J. (1996). Effects of phospholipase C inhibitors on $Ca^{2+}$ channel stimulation and $Ca^{2+}$ release from intracellular stores evoked by alpha 1A- and alpha 2A-adrenoceptors in rat portal vein myocytes. Biochem. Biophys. Res. Commun. 218, 30-34 https://doi.org/10.1006/bbrc.1996.0006
  25. McGehee, D.S., Aldersberg, M., Liu, K.P., Hsuing, S.C., Heath, M.J.S., and Tamir, H. (1997). Mechanism of extracellular $Ca^{2+}$receptor-stimulated hormone release from sheep thyroid parafollicular cell. J. Physiol. 502, 31-34 https://doi.org/10.1111/j.1469-7793.1997.031bl.x
  26. Monick, M.M., Carter, A.B., Gudmundsson, G., Mallampalli, R., Powers, L.S., and Hunninghake, G.W. (1999). A phosphatidylcholine-specific phospholipase C regulates activation of p42/44 mitogen-activated protein kinases in lipopolysaccharide-stimulated human alveolar macrophages. J. Immunol. 162, 3005-3012
  27. Muller-Decker, K. (1989). Interruption of TPA-induced signals by an antiviral and antitumoral xanthate compound: inhibition of a phospholipase C-type reaction. Biochem. Biophys. Res. Commun. 162, 198-205 https://doi.org/10.1016/0006-291X(89)91981-5
  28. Nofer, J.R., Junker, R., Seedorf, U., Assmann, G., Zidek, W., and Tepel, M. (2000). D609-phosphatidylcholine-specific phospholipase C inhibitor attenuates thapsigargin-induced sodium influx in human lymphocytes. Cell. Signal. 12, 289-296 https://doi.org/10.1016/S0898-6568(00)00068-1
  29. O'Flaherty, J.T., Chadwell, B.A., Kearns, M.W., Sergeant, S., and Daniel, L.W. (2001). Protein kinases C translocation responses to low concentrations of arachidonic acid. J. Biol. Chem. 276, 24743-24750 https://doi.org/10.1074/jbc.M101093200
  30. Pastorino, J.G., Simbula, G., Yamamoto, K., Glascott, P.A., Rothman, R.J., and Farber, J.L. (1996). The cytotoxicity of tumor necrosis factor depends on induction of the mitochondrial permeability transition. J. Biol. Chem. 271, 29792-29798 https://doi.org/10.1074/jbc.271.47.29792
  31. Pessin, M.S., and Raben, D.M., (1989). Molecular species analysis of 1,2-diglycerides stimulated by alpha-thrombin in cultured fibroblasts. J. Biol. Chem. 264, 8729-8738
  32. Pettitt, T.R., Martin, A., Horton, T., Liossis, C., Lord, J.M., and Wakelam, M.J. (1997). Diacylglycerol and phosphatidate generated by phospholipases C and D, respectively, have distinct fatty acid compositions and functions. Phospholipase D-derived diacylglycerol does not activate protein kinase C in porcine aortic endothelial cells. J. Biol. Chem. 272, 17354-17359 https://doi.org/10.1074/jbc.272.28.17354
  33. Ramoni, C., Spadaro, F., Barletta, B., Dupuis, M.L., and Podo, F. (2004). Phosphatidylcholine-specific phospholipase C in mitogen-stimulated fibroblasts. Exp. Cell. Res. 299, 370-82 https://doi.org/10.1016/j.yexcr.2004.05.037
  34. Ross, B.M., Kim, D.K., Bonventre, J.V., and Kish, S.J. (1995). Characterization of a novel phospholipase $A_2$ activity in human brain. J. Neurochem. 64, 2213-2221 https://doi.org/10.1046/j.1471-4159.1995.64052213.x
  35. Schutze, S., Potthoff, K., Machleidt, T., Berkovic, D., Wiegmann, K., and Kronke, M. (1992). TNF activates NF-kappa B by phosphatidylcholine-specific phospholipase C-induced "acidic" sphingomyelin breakdown. Cell 71, 765-776 https://doi.org/10.1016/0092-8674(92)90553-O
  36. Shin, K.J., Chung, C., Hwang, Y.A., Kim, S.H., Han, M.S., Ryu, S.H., and Suh, P.G. (2002). Phospholipase $A_2$-mediated $Ca^{2+}$ influx by 2,2',4,6-tetrachlorobiphenyl in PC12 cells. Toxicol. Appl. Pharmacol. 178, 37-43 https://doi.org/10.1006/taap.2001.9317
  37. Temes, E., Martin-Puig, S., Aragones, J., Jones, D.R., Olmos, G., Merida, I., and Landazuri, M.O. (2004). Role of diacylglycerol induced by hypoxia in the regulation of HIF-1alpha activity. Biochem. Biophys. Res. Commun. 315, 44-50 https://doi.org/10.1016/j.bbrc.2004.01.015
  38. Tschaikowsky, K., Schmidt, J., and Meisner, M., (1998). Modulation of mouse endotoxin shock by inhibition of phosphatidylcholinespecific phospholipase C. J. Pharmacol. Exp. Ther. 285, 800-804
  39. Wiegmann, K., Schutze, S., Machleidt, T., Witte, D., and Kronke, M., (1994). Functional dichotomy of neutral and acidic sphingomyelinases in tumor necrosis factor signaling. Cell 78, 1005-1015 https://doi.org/10.1016/0092-8674(94)90275-5
  40. Woo, C.H., Eom, Y.W., Yoo, M.H., You, H.J., Han, H.J., Song, W.K., Yoo, Y.J., Chun, J.S., and Kim, J.H. (2000). Tumor necrosis factor- alpha generates reactive oxygen species via a cytosolic phospholipase $A_2$-linked cascade, J. Biol. Chem. 275, 32357-32362 https://doi.org/10.1074/jbc.M005638200
  41. Xu, J., Weng, Y.I., Simonyi, A., Krugh, B.W., Liao, Z., Weisman, G.A., and Sun, G.Y. (2002). Role of PKC and MAPK in cytosolic $PLA_2$ phosphorylation and arachadonic acid release in primary murine astrocytes. J. Neurochem. 83, 259-270 https://doi.org/10.1046/j.1471-4159.2002.01145.x
  42. You, H.J., Seo, J.M., Moon, J.Y., Han, S.S., Ko, Y.G., and Kim, J.H. (2007). Leukotriene synthesis in response to $A_2$3187 is inhibited by methyl- beta-cyclodextrin in RBL-2H3 cells. Mol. Cells 23, 57-63
  43. Zamorano, J., Rivas, M.D., Garcia-Trinidad, A., Qu, C.K., and Keegan, A.D. (2003). Phosphatidylcholine-specific phospholipase C activity is necessary for the activation of STAT6. J. Immunol. 171, 4203-4209 https://doi.org/10.4049/jimmunol.171.8.4203
  44. Zhang, F., Zhao, G., and Dong, Z. (2001). Phosphatidylcholinespecific phospholipase C regulates activation of RAW264.7 macrophage-like cells by lipopeptide JBT3002. J. Leukoc. Biol. 69, 1060-1066
  45. Zhao, S., Du, X.Y., Chai, M.Q., Chen, J.S., Zhou, Y.C., and Song, J.G. (2002). Secretory phospholipase $A_2$ induces apoptosis via a mechanism involving ceramide generation. Biochim. Biophys. Acta 1581, 75-88 https://doi.org/10.1016/S1388-1981(02)00122-1