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Tyrosine Phosphorylation of Moesin in Arachidonic Acid–Stimulated Human Platelets

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Abstract

Moesin, a member of the ezrin/radixin/moesin (ERM) family of cytoskeletal proteins, has been implicated in dynamic membrane-based processes such as the formation and stabilization of filopodia. Ezrin is known to be a substrate of tyrosine kinases in activated T cells and epithelial growth factor–stimulated A431 cells. For the closely related 77-kD protein moesin, which shares 72% identity with ezrin on the basis of their amino acid sequences, a reversible phosphorylation on tyrosine residues has not yet been described. Because our scanning electron microscopy studies revealed the appearance of multiple, up to 3 μm long filopodia on the surface of activated human platelets, we investigated the participation of moesin in dynamic shape changes on platelet stimulation with arachidonic acid. Antimoesin immunoprecipitates obtained under denaturing conditions from lysates of resting platelets contained only low amounts of tyrosine-phosphorylated moesin. In lysates of arachidonic acid–stimulated platelets, the level of tyrosine phosphorylation was significantly increased. This activation-dependent phosphorylation of moesin was verified by probing antiphosphotyrosine immunoprecipitates from unstimulated and stimulated platelets with antimoesin antibodies. Tyrosine-phosphorylated moesin was detectable only in the presence of the tyrosine phosphatase inhibitor vanadate, suggesting that a coordinated balance between kinase and phosphatase activities controls the steady-state level of moesin phosphorylation.

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References

  1. Escolar G, Krumwiede M, White JG. Organization of the actin cytoskeleton of resting and activated platelets in suspension. Am J Pathol 1986;123:86–94.

    PubMed  Google Scholar 

  2. White JG. Arrangements of actin filaments in the cytoskeleton of human platelets. Am J Pathol 1984;117:207–217.

    PubMed  Google Scholar 

  3. Siess W, Siegel FL, Lapetina EG. Arachidonic acid stimulates the formation of 1,2-diacylglycerol and phosphatidic acid in human platelets. J Biol Chem 1983;258:11236–11242.

    PubMed  Google Scholar 

  4. 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.

    PubMed  Google Scholar 

  5. Lipfert L, Haimovich B, Schaller MD, Cobb BS, Parsons JT, Brugge JS. Integrin-dependent phosphorylation and activation of the protein tyrosine kinase pp125FAK in platelets. J Cell Biol 1992;119:905–912.

    PubMed  Google Scholar 

  6. Zhu B, O'Neill S, Saklatvala J, Tassi L, Mendelsohn ME. Phosphorylated HSP27 associates with the activation-dependent cytoskeleton in human platelets. Blood 1994; 84:3715–3723.

    PubMed  Google Scholar 

  7. Ozawa K, Kashiwada K, Takahashi M, Sobue K. Translocation of cortactin (p80/85) to the actin-based cytoskeleton during thrombin receptor-mediated platelet activation. Exp Cell Res 1995;221:197–204.

    PubMed  Google Scholar 

  8. Jackson SP, Schoenwaelder SM, Yuan Y, Rabinowitz I, Salem HH, Mitchell CA. Adhesion receptor activation of phosphatidylinositol 3-kinase. J Biol Chem 1994;269: 27093–27099.

    PubMed  Google Scholar 

  9. Golden A, Brugge JS. Thrombin treatment induces rapid changes in tyrosine phosphorylation in platelets. Proc Natl Acad Sci USA 1989;86:901–905.

    PubMed  Google Scholar 

  10. 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.

    PubMed  Google Scholar 

  11. Bertagnolli ME, Locke SJ, Hensler ME, Bray PF, Beckerle MC. Talin distribution and phosphorylation in thrombin-activated platelets. J Cell Sci 1993;106:1189–1199.

    PubMed  Google Scholar 

  12. Hagmann J, Burger MM. Phosphorylation of vinculin in human platelets spreading on a solid surface. J Cell Biochem 1992;50:237–244.

    PubMed  Google Scholar 

  13. Carroll RC, Gerrard JM. Phosphorylation of platelet actinbinding protein during platelet activation. Blood 1982;59: 466–471.

    PubMed  Google Scholar 

  14. Takeuchi K, Sato N, Kasahara H, et al. Perturbation of cell adhesion and microvilli formation by antisense oligonucleotides to ERM family members. J Cell Biol 1994; 125:1371–1384.

    PubMed  Google Scholar 

  15. Amieva MR, Furthmayr H. Subcellular localization of moesin in dynamic filopodia, retraction fibers, and other structures involved in substrate exploration, attachment, and cell-cell contacts. Exp Cell Res 1995;219:180–196.

    PubMed  Google Scholar 

  16. Turunen O, Wahlström T, Vaheri A. Ezrin has a COOH-terminal actin-binding site that is conserved in the ezrin protein family. J Cell Biol 1994;126:1445–1453.

    PubMed  Google Scholar 

  17. Tsukita S, Oishi K, Sato N, Sagara J, Kawai A, Tsukita S. ERM family members as molecular linkers between the cell surface glycoprotein CD44 and actin-based cytoskeletons. J Cell Biol 1994;126:391–401.

    PubMed  Google Scholar 

  18. Sato N, Funayama N, Nagafuchi A, Yonemura S, Tsukita S, Tsukita S. A gene family consisting of ezrin, radixin and moesin. Its specific localization at actin filament/plasma membrane association sites. J Cell Sci 1992;103:131–143.

    PubMed  Google Scholar 

  19. Franck Z, Gary R, Bretscher A. Moesin, like ezrin, colocalizes with actin in the cortical cytoskeleton in cultured cells, but its expression is more variable. J Cell Sci 1993; 105:219–231.

    PubMed  Google Scholar 

  20. Lankes WT, Furthmayr H. Moesin: Amember of the protein 4.1-talin-ezrin family of proteins. Proc Natl Acad Sci USA 1991;88:8297–8301.

    PubMed  Google Scholar 

  21. Nakamura F, Amieva MR, Furthmayr H. Phosphorylation of threonine 558 in the carboxyl-terminal actin-binding domain of moesin by thrombin activation of human platelets. J Biol Chem 1995;270:31377–31385.

    PubMed  Google Scholar 

  22. Takeuchi K, Kawashima A, Nagafuchi A, Tsukita S. Structural diversity of band 4.1 superfamily members. J Cell Sci 1994;107:1921–1928.

    PubMed  Google Scholar 

  23. Gary R, Bretscher A. Heterotypic and homotypic associations between ezrin and moesin, two putative membrane-cytoskeletal linking proteins. Proc Natl Acad Sci USA 1993; 90:10846–10850.

    PubMed  Google Scholar 

  24. Egerton M, Burgess WH, Chen D, Druker BJ, Bretscher A, Samelson LE. Identification of ezrin as an 81-kDa tyrosinephosphorylated protein in T cells. J Immunol 1992; 149:1847–1852.

    PubMed  Google Scholar 

  25. Helander TS, Carpen O, Turunen O, Kovanen PE, Vaheri A, Timonen T. ICAM-2 redistributed by ezrin as a target for killer cells. Nature 1996;382:265–268.

    PubMed  Google Scholar 

  26. Andréoli C, Martin M, Le Borgne R, Reggio H, Mangeat P. Ezrin has properties to self-associate at the plasma membrane. J Cell Sci 1994;107:2509–2521.

    PubMed  Google Scholar 

  27. Gary R, Bretscher A. Ezrin self-association involves binding of an N-terminal domain to a normally masked C-terminal domain that includes the F-actin binding site. Mol Biol Cell 1995;6:1061–1075.

    PubMed  Google Scholar 

  28. Fazioli F, Wong WT, Ullrich SJ, Sakaguchi K, Appella E, Di Fiore PP. The ezrin-like family of tyrosine kinase substrates: Receptor-specific pattern of tyrosine phosphorylation and relationship to malignant transformation. Oncogene 1993;8:1335–1345.

    PubMed  Google Scholar 

  29. Bretscher A. Rapid phosphorylation and reorganization of ezrin and spectrin accompany morphological changes induced in A-431 cells by epidermal growth factor. J Cell Biol 1989;108:921–930.

    PubMed  Google Scholar 

  30. Martin M, Andréoli C, Sahuquet A, Montcourrier P, Algrain M, Mangeat P. Ezrin NH2-terminal domain inhibits the cell extension activity of the COOH-terminal domain. J Cell Biol 1995;128:1081–1093.

    PubMed  Google Scholar 

  31. Gould KL, Cooper JA, Bretscher A, Hunter T. The proteintyrosine kinase substrate, p81, is homologous to a chicken microvillar core protein. J Cell Biol 1986;102:660–669.

    PubMed  Google Scholar 

  32. Krieg J, Hunter T. Identification of the twomajor epidermal growth factor-induced tyrosine phosphorylation sites in the microvillar core protein ezrin. J Biol Chem 1992;267: 19258–19265.

    PubMed  Google Scholar 

  33. Bhattacharya S, Fu C, Bhattacharya J, Greenberg S. Soluble ligands of the αvβ3 integrin mediate enhanced tyrosine phosphorylation of multiple proteins in adherent bovine pulmonary artery endothelial cells. J Biol Chem 1995; 270:16781–16787.

    PubMed  Google Scholar 

  34. Chen J, Cohn JA, Mandel LJ. Dephosphorylation of ezrin as an early event in renal microvillar breakdown and anoxic injury. Proc Natl Acad Sci USA 1995;92:7495–7499.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Cardiology, University of Göttingen, Göttingen, Germany

    Thomas Meyer, Thomas Uher & Arnd B. Buchwald

  2. Institute of Anatomy, University of Göttingen, Göttingen, Germany

    Peter Schwartz

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  1. Thomas Meyer
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  2. Thomas Uher
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  4. Arnd B. Buchwald
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Meyer, T., Uher, T., Schwartz, P. et al. Tyrosine Phosphorylation of Moesin in Arachidonic Acid–Stimulated Human Platelets. J Thromb Thrombolysis 6, 117–124 (1998). https://doi.org/10.1023/A:1008845421381

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