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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)

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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
    CrossrefPubMedSearch in Google Scholar
  • 2 Ferrell JE, Martin GS. Platelet tyrosine-specific protein phosphorylation is regulated by thrombin. Mol Cell Biol 1988; 8: 3603-3610
    CrossrefPubMedSearch in Google Scholar
  • 3 Golden A, Brugge JS. Thrombin treatment induces rapid changes in tyrosine phosphorylation in platelets. Proc Natl Acad Sci (USA) 1989; 86: 901-905
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 14 Rhee SG, Choi KD. Regulation of inositol phospholipid-specific phospholipase C isozymes. J Biol Chem 1992; 267: 12393-12396
    PubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 19 Nozawa Y, Nakashima S, Nagata K. Phospholipid-mediated signalling in receptor activation of platelets. Biochim Biophys Acta 1991; 1082: 219-238
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 22 Pollock WK, Armstrong RA, Brydon LJ, Jones RL, MacIntyre DE. Thromboxane-induced phosphatidate formation in human platelets. Biochem J 1984; 219: 833-842
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 25 Waite M. The Phospholipases. In: Handbook of Lipid Research, Vol. 5. Ha-nahan DJ, ed. New York: Plenum Press 1987, pp 111-113
    PubMed
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 31 Blenis J. Signal transduction via the MAP kinases: proceed at your own RSK. Proc Natl Acad Sci (USA) 1993; 90: 5889-5892
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 33 McNicol A, Roberston C, Gerrard JM. Vanadate activates platelets by enhancing arachidonic acid release. Blood 1993; 81: 2329-2338
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    PubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 42 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedSearch in Google Scholar
  • 43 Farndale RW, Winkler AB, Martin BR, Barnes MJ. Inhibition of human platelet adenylate cyclase by collagen fibres. Biochem J 1992; 282: 25-32
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 51 Furman MI, Gardner TM, Goldschmidt-Clermont PJ. Mechanisms of cyto-skeletal reorganization during platelet activation. Thromb Haemost 1993; 70: 229-232
    Thieme ConnectPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 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
    CrossrefPubMedSearch in Google Scholar
  • 54 Shattil SJ, Brugge JS. Protein tyrosine phosphorylation and the adhesive functions of platelets. Curr Opin in Cell Biol 1991; 3: 869-879
    PubMedSearch in Google Scholar