Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effects of Protein Kinase G on Phospholipase D Activity of Human Neutrophils

호중구에서 phospholipase D의 활성에 대한 protein kinase G의 영향

  • 박지연 (동아대학교 의과대학 생화학교실) ;
  • 이민정 (동아대학교 암분자치료연구센터) ;
  • 장민정 (동아대학교 암분자치료연구센터) ;
  • 이선영 (동아대학교 의과대학 생화학교실) ;
  • 배외식 (동아대학교 의과대학 생화학교실) ;
  • 곽종영 (동아대학교 암분자치료연구센터)
  • Published : 2003.12.01
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Abstract

Phospholipase D (PLD) plays an important role as a signaling molecule in the activation of neutrophils. In this study, effect of nitric oxide (NO) and cGMP on the activation of PLD in human neutrophils was investigated. Sodium nitroprusside (SNP), an agent to produce NO spontaneously in cells, alone increased PLD activity and the maximal activation was obtained with 0.5 mM SNP. Dibutyryl-cAMP, an agent to increase an intracellular cAMP concentration inhibited formyl-Met-Leu-Phe (fMLP)-stimulated PLD activity but 8-bromo-cGMP (300 $\mu$M), an agent to increase an intracellular cGMP concentration did not affect basal and fMLP-stimulated PLD activity. NO-induced activation of PLD was not blocked by KT 5823, an inhibitor of cGMP-dependent protein kinase (PKG), suggesting that NO-induced PLD activation is not mediated by cGMP. NO also stimulated p38 mitogen activated protein kinase (MAPK) in human neutrophils, indicated by increased phosphorylation of p38 MAPK in Western blotting. NO-induced phosphorylation of p38 MAPK was not inhibited by KT 5823 or n-butanol. RhoA, an regulatory factor of PLD activation was trans-located from cytosolic fraction to plasma membranes by fMLP or phorbol ester, and fMLP-stimulated but not phorbol ester-stimulated translocation of RhoA was inhibited by cGMP. These results suggest that NO stimulates PLD activity through other unidentified facto.(s) than cGMP even though cGMP inhibits the artivation of RhoA.

Phosphipase D(PLD)는 호중구의 활성에서 중요한 신호전달 인자로 작용한다. 본 연구에서는 호중구에서 PLD의 활성화에 대한 nitric oxide(NO)와 cGMP의 영향을 조사하였다. 세포 내 NO의 생성을 증가시키는 물질인 sodium nitroprusside (SNP)를 단독으로 처리하였을 때 SNP를 처리하지 않은 세포에 비교하여 PLD 활성은 0.5 mM 농도에서 2배 이상 증가하였다. 세포 내 cAMP의 농도를 증가시키는 물질인 dibutyryl-cAMP를 처리하였을 때 formyl-Met-Leu-Phe(fMLP)에 의한 PLD활성은 억제되었으나 cGMP를 증가시키는 물질인 8-bromo-cGMP(300 $\mu$M)를 단독으로나 fMLP와 같이 처리하였을 때 PLD의 활성은 큰 영향이 없었다. NO에 의한 PLD의 활성은 cGMP-의존형 인산화 효소인 protein kinase G(PKG)의 억제제인 KT 5823에 의하여 억제되지 않았는데 이러한 결과는 PKG 이외의 경로를 통하여 일어남을 제시한다. NO를 처리한 호중구에서 p38 mitogen activated protein kinase(MAPK)가 활성화되어 인산화된 p38 MAPK가 Western blot에서 증가되었다. NO에 의한 p38 MAPK의 인산화는 KT 5823에 의하여 억제되지 않았고 PLD 억제제인 n-butanol에 의하여도 영향을 받지 않았다. PLD 활성의 인자인 RhoA는 fMLP나 phorbol myristate acetate(PMA)의 자극에 의하여 세포질로부터 세포막으로 전이가 되었으나 cGMP의 전처리에 의하여 fMLP에 의한 RhoA의 전이는 억제되었으나 PMA에 의한 전이는 영향을 받지 않았다. 이들 결과들은 호중구 내 증가된 cGMP가 RhoA를 억제하였으나 세포 내 증가된 NO는 cGMP 이외의 인자를 통하여 PLD의 활성화를 일으킨다는 것을 제시하고 있다.

Keywords

References

  1. D. J. Biol. Chem. v.273 Determination of interaction sites on the small G protein RhoA for phospholipase Bae,C.D.;D.S.Min;I.N.Fleming;J.H.Exton
  2. Am. J. Physiol. v.268 cGMP antagonizes angiotensin-mediated phosphatidylcholine hydrolysis and C kinase activation in mesangial cells Barnett,R.L.;L.Ruffini;L.Ramsammy;R.Pasmantier;M.M.Friedlaender;E.P.Nord
  3. J. Biol. Chem. v.276 Phospholipase D is required in the signaling pathway leading to p38 MAPK activation in neutrophil-like HL-60 cells, stimulated by N-formyl-methionyl-leucyl-phenylalanine Bechoua,S.;L.W.Daniel https://doi.org/10.1074/jbc.M101265200
  4. J. Biol. Chem. v.264 Phosphatidylcholine hydrolysis by phospholipase D determines phosphatidate and diacylglyceride lavels in chemotactic peptidestimulated human neutrophils. Involvement of phosphatidate phosphohydrolase in signal transduction Billah,M.M.;S.Eckel;T.J.Mullmann;R.W.Egan;M.I.Siegel
  5. J. Biol. Chem. v.274 Activation of p38 mitogen-activated protein kinase by lipopolysaccharide in human neutrophils requires nitric oxide-dependent cGMP accumulation Browning,D.D.;N.D.Windes;R.D.Ye https://doi.org/10.1074/jbc.274.1.537
  6. J. Biol. Chem. v.275 Nitric oxide activation of p38 mitogen-activated protein kinase in 293T fibroblasts requires cGMP-dependent protein kinase Browning,D.D.;M.P.McShane;C.Marty;R.D.Ye https://doi.org/10.1074/jbc.275.4.2811
  7. J. Biol. Chem. v.273 Protein kinase C eta mediates lipopolysaccharide-induced nitric-oxide synthase expression in primary astrocytes Chen,C.C.;J.K.Wang;W.C.Chen;S.B.Lin https://doi.org/10.1074/jbc.273.31.19424
  8. Endocrinology v.141 Interleukin-1 β regulates phospholipase D-1 expression in rat pancreatic β-cells Chen,M.C.;V.Paez-Espinosa;N.Welsh;D.L.Eizirik https://doi.org/10.1210/en.141.8.2822
  9. Biochim. Biophys. Acta v.1113 G-protein-regulated phospholipase C, D and A2-mediated signalling in neutrophils Cockcroft,S. https://doi.org/10.1016/0304-4157(92)90036-A
  10. J. Infect. Dis. v.177 Effects of nitric oxide on chemotaxis and endotoxin-induced interleukin-8 production in human neutrophils Corriveau,C.C.;P.J.Madara;A.L. Van Dervort;M.M.Tropea;R.A.Wesley;R.L.Danner https://doi.org/10.1086/513829
  11. Biochim. Biophys. Acta v.1439 Regulation of phospholipase D Exton,J.H. https://doi.org/10.1016/S1388-1981(99)00089-X
  12. Annu. Rev. Physiol. v.56 Structure and function of cyclic nucleotide-dependent protein kinases Francis,S.H.;J.D.Corbin https://doi.org/10.1146/annurev.ph.56.030194.001321
  13. J. Cell Sci. v.113 Rising behind NO: cGMP-dependent protein kinases Hofmann,F.;A.Ammendola;J.Schlossmann
  14. FASEB J. v.7 Nitric oxide synthesis inhibition induces leukocyte adhesion via superoxide and mast cells Kubes,P.;S.Kanwar;X.F.Niu;J.P.Gaboury
  15. J. Biol. Chem. v.270 Rho A and a cytosolic 50-kDa factor reconstitute GTP γS-dependent phospholipase D activity in human neutrophil subcellular fractions Kwak,J.Y.;I.Lopez;D.J.Uhlinger;S.H.Ryu;J.D.Lambeth https://doi.org/10.1074/jbc.270.45.27093
  16. Biochem. Biophys. Res. Commun. v.267 Downregulation of Phospholipase D by Protein Kinase A in a Cell-Free System of Human Neutrophils Kwak,J.Y.;D.J.Uhlinger https://doi.org/10.1006/bbrc.1999.1941
  17. Recent Research Development in Biophysical Chemistry v.2 Regulation of phospholipase D activity in Human neutrophils by RhoA and cAMP-dependent protein kinase Kwak,J.Y.;D.J.Uhlinger;Condat,C.(ed.);A.Baruzzi(ed.)
  18. EMBO J. v.15 Protein kinase A phosphorylation of RhoA mediates the morphological and functional effects of cyclic AMP in cytotoxic lymphocytes Lang,P.;F.Gesbert;M.Delespine-Carmagnat;R.Stancou;M.Pouchelet;J.Bertoglio
  19. Nature v.364 A redox-based mechanism for the neuroprotective and neurodestructive effects of nitric oxide and related nitroso-compounds Lipton,S.A.;Y.B.Choi;Z.H.Pan;S.Z.Lei;H.S.Chen;N.J.Sucher;J.Loscalzo;D.J.Singel;J.S.Stamler https://doi.org/10.1038/364626a0
  20. FEBS Lett. v.413 Nitric oxide inhibits enterocyte mitochondrial phospholipase D. Madesh,M.;K.A.Balasubramanian https://doi.org/10.1016/S0014-5793(97)00903-4
  21. Nature v.323 Growth factor-like action of phosphatidic acid Moolenaar,W.H.;W.Kruijer;B.C.Tilly;I.Veriaan;A.J.Biermann;S.W.deLaat https://doi.org/10.1038/323171a0
  22. Biochemistry v.34 Inhibition of ribonucleotide reductase by nitric oxide derived from thionitrites: reversible modifications of both subunits Roy,B.;M.Lepoivre;Y.Henry;M.Fontecave https://doi.org/10.1021/bi00016a012
  23. Biochem. Biophys. Res. Commun. v.280 cGMP-Dependent protein kinase phosphorylates and inactivates RhoA Sawada,N.;H.Itoh;J.Yamashita;K.Doi;M.Inoue;K.Masatsugu;Y.Fukunaga;S.Sakaguchi;M.Sone;K.Yamahara;T.Yurugi;K.Nakao https://doi.org/10.1006/bbrc.2000.4194
  24. J. Biol. Chem. v.266 Cyclic AMP-elevating agents block chemoattractant activation of diradylglycerol generation by inhibiting phospholipase D activation Tyagi,S.R.;S.C.Olson;D.N.Burnham;J.D.Lambeth
  25. Biochem. Pharmacol. v.62 Inhibition of extracellular $Ca ^{2+}$ entry by YC-1, an activator of soluble guanylyl cyclase, through a cyclic GMP-independent pathway in rat neutrophils Wang,J.P.;L.C.Chang;L.J.Huang;S.C.Kuo https://doi.org/10.1016/S0006-2952(01)00725-0
  26. Biochem. Pharmacol. v.63 Inhibition of superoxide anion generation by YC-1 in rat neutrophils through cyclic GMP-dependent and -independent mechanisms Wang,J.P.;L.C.Chang;S.L.Raung;M.F.Hsu;L.J.Huang;S.C.Kuo https://doi.org/10.1016/S0006-2952(01)00882-6
  27. Br. J. Pharmacol. v.122 Investigation of the role of nitric oxide and cyclic GMP in both the activation and inhibition of human neutrophils Wanikiat,P.;D.F.Woodward;R.A.Armstrong https://doi.org/10.1038/sj.bjp.0701477
  28. Am. J. Physiol. v.265 Regulation of human neutrophil degranulation by LY-83583 and L-arginine: role of cGMP-dependent protein kinase Wyatt,T.A.;T.M.Lincoln;K.B.Pryzwansky
  29. J. Biol. Chem. v.274 Interaction of the small G protein RhoA with the C terminus of human phospholipase D1 Yamazaki,M.;Y.Zhang;H.Watanabe;T.Yokozeki;S.Ohno;K.Kaibuchi;H.Shibata;H.Mukai;Y.Ono;M.A.Frohman;Y.Kanaho https://doi.org/10.1074/jbc.274.10.6035
  30. Cell v.52 Cellular ras activity and phospholipid metabolism Yu,C.L.;M.H.Tsai;D.W.Stacey https://doi.org/10.1016/0092-8674(88)90531-4