Thromb Haemost 1994; 72(04): 634-642
DOI: 10.1055/s-0038-1648927
Original Article
Schattauer GmbH Stuttgart

The Tyrosine Kinase Inhibitors, Genistein and Methyl 2,5-Dihydroxycinnamate, Inhibit the Release of (3H)Arachidonate from Human Platelets Stimulated by Thrombin or Collagen

Philip G Hargreaves
1   The Dept. of Biochemistry, University of Cambridge, Cambridge, UK
,
Ellen F Licking
1   The Dept. of Biochemistry, University of Cambridge, Cambridge, UK
,
Paul Sargeant
2   The Physiological Laboratory, University of Cambridge, Cambridge, UK
,
Stewart O Sage
2   The Physiological Laboratory, University of Cambridge, Cambridge, UK
,
Michael J Barnes
3   Strangeways Research Laboratory, Cambridge, UK
,
Richard W Farndale
1   The Dept. of Biochemistry, University of Cambridge, Cambridge, UK
› Author Affiliations
Further Information

Publication History

Received: 10 May 1994

Accepted after revision23 June 1994

Publication Date:
06 July 2018 (online)

Summary

We have investigated the effects of the tyrosine kinase inhibitors, genistein and methyl 2,5-dihydroxycinnamate, on [3H]arachidonic acid release from human platelets. Both tyrosine kinase inhibitors blocked, in a dose-dependent manner, the release of arachidonic acid stimulated by thrombin or suspensions of collagen fibres. Blockade by the tyrosine kinase inhibitors occurred early in the arachidonate release time course. Both genistein and methyl 2,5-dihydroxycinnamate also inhibited tyrosine phosphorylation in platelets. The inhibitors were specific in that they did not affect protein kinase C activity, ATP levels or mobilization of Ca2+ from internal stores. These findings suggest a role for tyrosine kinase activity in the regulation of phospholipase A2 in platelets stimulated by the physiological ligands, thrombin and collagen

 
  • References

  • 1 Siess W. Molecular mechanisms of platelet activation. Physiol Rev 1989; 69: 58-178
  • 2 Ferrell JE, Martin GS. Platelet tyrosine-specific protein phosphorylation is regulated by thrombin. Mol Cell Biol 1988; 8: 3603-3610
  • 3 Golden A, Brugge JS. Thrombin treatment induces rapid changes in tyrosine phosphorylation in platelets. Proc Natl Acad Sci (USA) 1989; 86: 901-905
  • 4 Huang M-M, Bolen JB, Barnwell JW, Shattil SJ, Brugge JS. Membrane glycoprotein IV (CD36) is physically associated with the Fyn, Lyn and Yes protein-tyrosine kinases in human platelets. Proc Natl Acad Sci (USA) 1991; 88: 7844-7848
  • 5 Lipfert L, Haimovich B, Schaller MD, Cobb BS, Parsons JT, Brugge JS. In-tegrin-dependent phosphorylation and activation of the protein tyrosine kinase ppl25fak in platelets. J Cell Biol 1992; 119: 905-912
  • 6 Taniguchi T, Kitagawa H, Yasue S, Yanagi S, Sakai K, Asahi M, Ohta S, Takeuchi F, Nakamura S, Yamamura H. Protein-tyrosine kinase p72(syk) is activated by thrombin and is negatively regulated through Ca2+ mobilization in platelets. J Biol Chem 1993; 268: 2277-2279
  • 7 Nakamura S, Yamamura H. Thrombin and collagen induce rapid phosphorylation of a common set of cellular proteins on tyrosine in human platelets. J Biol Chem 1989; 264: 7089-7091
  • 8 Haimovich B, Lipfert L, Brugge JS, Shattil SJ. Tyrosine phosphorylation and cytoskeletal reorganization in platelets are triggered by interaction of integrin receptors with their immobilized ligands. J Biol Chem 1993; 268: 15868-15877
  • 9 Polanowska-Grabowska R, Geanacopoulos M, Gear ARL. Platelet adhesion to collagen via the a2(3, integrin under arterial flow conditions causes rapid tyrosine phosphorylation of ppl25fak. Biochem J 1993; 296: 543-547
  • 10 Salari H, Duronio V, Howard SL, Demos M, Jones K, Reany A, Hudson AT, Pelech SL. Erbstatin blocks platelet activating factor-induced protein-tyrosine phosphorylation, polyphosphoinositide hydrolysis, protein kinase C activation, serotonin secretion and aggregation of rabbit platelets. FEBS Lett 1990; 263: 104-108
  • 11 Rendu F, Eldor A, Grelac F, Levy-Toledano S, Levitzki A. Tyrosine kinase blockers: new platelet activation inhibitors. Blood Coagulation and Fibrinolysis 1990; 1: 713-716
  • 12 Bachelot C, Cano E, Grelac F, Saleun S, Druker BJ, Levy-Toledano S, Fischer S, Rendu F. Functional implications of tyrosine phosphorylation in platelets. Simultaneous studies with different agonists and inhibitors. Biochem J 1992; 284: 923-928
  • 13 Margolis B, Rhee SG, Felder S, Mervic M, Lyall R, Levitzki A, Ullrich A, Zilberstein A, Schlessinger J. EGF induces tyrosine phosphorylation of phospholipase C-II: a potential mechanism for EGF signalling. Cell 1989; 57: 1101-1107
  • 14 Rhee SG, Choi KD. Regulation of inositol phospholipid-specific phospholipase C isozymes. J Biol Chem 1992; 267: 12393-12396
  • 15 Vostal JG, Jackson WL, Shulman NR. Cytosolic and stored calcium antagonistically control tyrosine phosphorylation of specific platelet protein. J Biol Chem 1991; 266: 16911-16916
  • 16 Sargeant P, Farndale RW, Sage SO. The tyrosine kinase inhibitors methyl 2,5-dihydroxycinnamate and genistein reduce thrombin-evoked tyrosine phosphorylation and Ca2+ entry in human platelets. FEBS Lett 1993; 315: 242-246
  • 17 Sargeant P, Farndale RW, Sage SO. ADP- and thapsigargin-evoked Ca2+ entry and protein tyrosine phosphorylation are inhibited by the tyrosine kinase inhibitors genistein and methyl-2,5-dihydroxycinnamate in fura-2-loaded human platelets. J Biol Chem 1993; 268: 18151-18156
  • 18 Sargeant P, Farndale RW, Sage SO. Calcium store depletion in dimethyl BAPTA-loaded human platelets increases protein tyrosine phosphorylation in the absence of a rise in cytosolic calcium. Exp Physiol 1994; 79: 269-272
  • 19 Nozawa Y, Nakashima S, Nagata K. Phospholipid-mediated signalling in receptor activation of platelets. Biochim Biophys Acta 1991; 1082: 219-238
  • 20 Turini ME, Holub BJ. Eicosanoid/thromboxane A2-independent and -dependent generation of lysoplasmenylethanolamine via phospholipase A2 in collagen-stimulated human platelets. Biochem J 1993; 289: 641-646
  • 21 Purdon AD, Patelumas D, Smith JB. Evidence for the release of arachi-donic acid through the selective action of phospholipase A2 in thrombin-stimulated human platelets. Biochim Biophys Acta 1987; 920: 205-214
  • 22 Pollock WK, Armstrong RA, Brydon LJ, Jones RL, MacIntyre DE. Thromboxane-induced phosphatidate formation in human platelets. Biochem J 1984; 219: 833-842
  • 23 Kienast J, Arnout J, Pfliegler G, Deckmyn H, Hoet B, Vermylen J. Sodium fluoride mimics effects of both agonists and antagonists on intact human platelets by simultaneous modulation of phospholipase C and adenylate cyclase activity. Blood 1987; 69: 859-866
  • 24 Silk ST, Clejan S, Witkom K. Evidence of GTP-binding protein regulation of phospholipase A2 activity in isolated human platelet membrane. J Biol Chem 1989; 264: 21466-21469
  • 25 Waite M. The Phospholipases. In: Handbook of Lipid Research, Vol. 5. Ha-nahan DJ, ed. New York: Plenum Press 1987, pp 111-113
  • 26 Stasi M, Gresele P, Porcellati S, Quero E, Nenci GG, Goracci G. Activation of phospholipase A2 and (3-thromboglobulin release in human platelets: comparative effects of thrombin and fluoroaluminate stimulation. Biochim Biophys Acta 1992; 1124: 279-287
  • 27 Kramer RM, Roberts EF, Manetta J, Putnam JE. The Ca2+-sensitive cytosolic phospholipase A2 is a 100-kDa protein in human monoblast U937 cells. J Biol Chem 1991; 266: 5268-5272
  • 28 Kramer RM, Roberts EF, Manetta J, Hyslop PA, Jakubowski JA. Thrombin-induced phosphorylation and activation of Ca2+-sensitive cytosolic phospholipase A2 in human platelets. J Biol Chem 1993; 268: 26796-26804
  • 29 Lin L-L, Wartmann M, Lin AY, Knopf JL, Seth A, Davis RJ. cPLA2 is phosphorylated and activated by MAP kinase. Cell 1993; 72: 269-278
  • 30 Winitz S, Gupta S, Qian N-X, Heasley LE, Nemenoff RA, Johnson GL. Expression of mutant Gi2 a subunit inhibits ATP and thrombin stimulation of cytoplasmic phospholipase A2-mediated arachidonic acid release independent of Ca2+ and mitogen-activated protein kinase regulation. J Biol Chem 1994; 269: 1889-1895
  • 31 Blenis J. Signal transduction via the MAP kinases: proceed at your own RSK. Proc Natl Acad Sci (USA) 1993; 90: 5889-5892
  • 32 Nakashima S, Chatani Y, Nakamura M, Miyoshi N, Kohno M, Nozawa Y. Tyrosine phosphorylation and activation of mitogen-activated protein kinases by thrombin in human platelets: possible involvement in late arachidonic acid release. Biochem Biophys Res Commun 1994; 198: 497-503
  • 33 McNicol A, Roberston C, Gerrard JM. Vanadate activates platelets by enhancing arachidonic acid release. Blood 1993; 81: 2329-2338
  • 34 Zor U, Ferber E, Gergely P, Szucs K, Dombradi V, Goldman R. Reactive oxygen species mediate phorbol ester-regulated tyrosine phosphorylation and phospholipase A2 activation: potentiation by vanadate. Biochem J 1993; 295: 879-888
  • 35 Banga HS, Simons ER, Brass LF, Rittenhouse SE. Activation of phospholi-pases A and C in human platelets exposed to epinephrine: role of glycoprotein iib/iiia and dual role of epinephrine. Proc Natl Acad Sci (USA) 1986; 83: 9197-9201
  • 36 Nakano T, Terawaki A, Arita H. Signal transduction in collagen-stimulated rat platelets is composed of three stages. J Biochem 1987; 101: 1169-1180
  • 37 Akiyama T, Ishida J, Nakagawa S, Ogawara H, Watanabe S-I, Itoh N, Shi-buya M, Fukami Y. Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem 1987; 262: 5592-5595
  • 38 Asahi M, Yanagi S, Ohta S, Inazu T, Sakai K, Takeuchi F, Taniguchi T, Ya-mamura H. Thrombin-induced platelet aggregation is inhibited by protein-tyrosine kinase inhibitors, ST638 and genistein. FEBS Lett 1992; 309: 10-14
  • 39 Isshiki K, Imoto M, Sawa T, Takeuchi F, Umezawa H. Inhibition of tyrosine protein kinase by synthetic erbstatin analogs. J Antibiot 1987; 40: 1209-1210
  • 40 Umezawa K, Hori T, Tajima H, Imoto M, Isshiki K, Takeuchi T. Inhibition of epidermal growth factor-induced DNA synthesis by tyrosine kinase inhibitors. FEBS Lett 1990; 260: 198-200
  • 41 Joseph S, Krishnamurthi S, Kakkar VV. Differential effects of the diacyl-glycerol kinase inhibitor R59022 on thrombin versus collagen-induced human platelet secretion. Biochim Biophys Acta 1988; 969: 9-17
  • 42 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970; 227: 680-685
  • 43 Farndale RW, Winkler AB, Martin BR, Barnes MJ. Inhibition of human platelet adenylate cyclase by collagen fibres. Biochem J 1992; 282: 25-32
  • 44 Smith JB, Selak MA, Dangelmaier C, Daniel JL. Cytosolic calcium as a second messenger for collagen-induced platelet responses. Biochem J 1992; 288: 925-929
  • 45 Smith JB, Dangelmaier C, Daniel JL. Elevation of cAMP in human platelets inhibits thrombin- but not collagen-induced tyrosine phosphorylation. Biochem Biophys Res Commun 1993; 191: 695-700
  • 46 Yaish P, Gazit A, Gilon C, Levitzki A. Blocking of EGF-dependent cell proliferation by EGF receptor kinase inhibitors. Science 1988; 242: 933-935
  • 47 Tyers M, Rachubinski RA, Stewart MI, Varrichio AM, Shorr RGL, Haslam RJ, Harley CB. Molecular cloning and expression of the major protein kinase C substrate of platelets. Nature 1988; 333: 470-473
  • 48 Oda A, Druker BJ, Smith M, Salzman EW. Inhibition by sodium nitroprus-side or PGE, of tyrosine phosphorylation induced in platelets by thrombin or ADP. Am J Physiol 1992; 262: C701-C707
  • 49 Pumiglia K, Huang C-K, Feinstein M. Elevation of cAMP but not cGMP inhibits thrombin-stimulated tyrosine phosphorylation in human platelets. Biochem Biophys Res Commun 1990; 171: 738-745
  • 50 Guinebault C, Payrastre B, Sultan C, Mauco G, Breton M, Levy-Toledano S, Plantavid M, Chap H. Tyrosine kinases and phosphoinositide metabolism in thrombin-stimulated human platelets. Biochem J 1993; 292: 851-856
  • 51 Furman MI, Gardner TM, Goldschmidt-Clermont PJ. Mechanisms of cyto-skeletal reorganization during platelet activation. Thromb Haemost 1993; 70: 229-232
  • 52 Samiei M, Sanghera JS, Pelech SL. Activation of myelin basic protein and S6 peptide kinases in phorbol ester-treated and PAF-treated sheep platelets. Biochim Biophys Acta 1993; 1176: 287-298
  • 53 Papkoff J, Chen R-H, Blenis J, Forsman J. p42 mitogen-activated protein kinase and p90 ribosomal S6 kinase are selectively phosphorylated and activated during thrombin-induced platelet activation and aggregation. Mol Cell Biol 1994; 14: 463-472
  • 54 Shattil SJ, Brugge JS. Protein tyrosine phosphorylation and the adhesive functions of platelets. Curr Opin in Cell Biol 1991; 3: 869-879