Subscribe to RSS
DOI: 10.1055/s-0037-1615073
Rap1b Association with the Platelet Cytoskeleton Occurs in the Absence of Glycoproteins IIb/IIIa
Publication History
Received
09 December 1997
Accepted after resubmission
19 November 1997
Publication Date:
07 December 2017 (online)
Summary
Platelet membrane glycoproteins (GP) IIb/IIIa and rap1b, a 21 kDa GTP binding protein, associate with the triton-insoluble, activation-dependent platelet cytoskeleton with similar rates and divalent cation requirement. To examine the possibility that GPIIb/IIIa was required for rap1b association with the cytoskeleton, experiments were performed to determine if the two proteins were linked under various conditions. Chromatography of lysates from resting platelets on Sephacryl S-300 showed that GPIIb/IIIa and rap1b were well separated and distinct proteins. Immunoprecipitation of GPIIb/IIIa from lysates of resting platelets did not produce rap1b or other low molecular weight GTP binding proteins and immunoprecipitation of rap1b from lysates of resting platelets did not produce GPIIb/IIIa. Finally, rap1b was associated with the activation-dependent cytoskeleton of platelets from a patient with Glanzmann’s thrombasthenia who lacks surface expressed glycoproteins IIb and IIIa. Based on these findings, we conclude that no association between GPIIb/IIIa and rap1b is found in resting platelets and that rap1b association with the activation-dependent cytoskeleton is at least partly independent of GPIIb/IIIa.
-
References
- 1 Fox JEB. The platelet cytoskeleton. Thromb Haemost 1993; 70: 884-93.
- 2 Burn P, Rotman A, Meyer RK, Burger MM. Diacylglycerol in large α-actinin/actin complexes and in the cytoskeleton of activated platelets. Nature 1985; 314: 469-72.
- 3 Burn P. Phosphatidylinositol cycle and its possible involvement in the regulation of cytoskeleton-membrane interactions. J Cell Biol 1988; 36: 15-24.
- 4 Nahas N, Plantavid M, Mauco G, Chap H. Association of phosphatidylinositol kinase and phosphatidylinositol 4-phosphate kinase activities with the cytoskeleton in human platelets. FEBS Letters 1989; 246: 30-4.
- 5 Grondin P, Plantavid M, Sultan C, Breton M, Mauco G, Chap H. Interaction of pp 60c-src, phospholipase C, inositol-lipid, and diacylglycerol kinases with the cytoskeletons of thrombin-stimulated platelets. J Biol Chem 1991; 266: 15705-9.
- 6 Zhang J, Fry MJ, Waterfield MD, Jaken S, Liao L, Fox JEB. and Ritten-house SE, Activated phosphoinositide 3-kinase associates with membrane skeleton in thrombin-exposed platelets. J Biol Chem 1992; 267: 4686-92.
- 7 Oda A, Druker BJ, Smith M, Salzman EW. Association of pp 60src with triton X-100 insoluble residue in human blood platelets requires platelet arregation and actin polymerization. J Biol Chem 1992; 267: 20075-81.
- 8 Fox JEB, Lipfert L, Clark EA, Reynolds CC, Austin CD, Brugge JS. On the role of the platelet membrane skeleton in mediating signal transduction. Association of GPIIb/IIIa, pp 60c-src, pp 62c-yes, and the p 21ras GTPase-activating protein with the membrane skeleton. J Biol Chem 1993; 268: 25973-84.
- 9 Pumiglia KM, Feinstein MB. Thrombin and thrombin receptor agonist peptide induce tyrosine phosphorylation and tyrosine kinases in the platelet cytoskeleton. Translocation of pp60(c-src) and integrin αIIbβ3 (glycoprotein IIb-IIIa) is not required for aggregation, but is dependent on formation of large aggregate structures. Biochem J 1993; 294: 253-60.
- 10 Li RY, Gaits F, Ragab A, Ragab-Thomas JMF, Chap H. Translocation of SH2-containing protein tyrosine phosphatase (SH-PTP1) to the cytoskeleton of thrombin-activated platelets. FEBS Letters 1994; 343: 89-93.
- 11 Phillips DR, Jennings LK, Edwards HH. Identification of membrane proteins mediating the interaction of human platelets. J Cell Biol 1980; 86: 77-86.
- 12 Painter RG, Prodouz KN, Gaarde W. Isolation of a subpopulation of glyco-protein IIb/IIIa from platelet membranes that is bound to membrane actin. J Cell Biol 1994; 100: 652-7.
- 13 Kouns WC, Fox CF, Lamoreaux WJ, Coons LB, Jennings LK. The effect of glycoprotein IIb-IIIa receptor occupancy on the cytoskeleton of resting and activated platelets. J Biol Chem 1991; 266: 13891-900.
- 14 Bertagnolli ME, Beckerle M. Evidence for the selective association of a subpopulation of GPIIb-IIIa with the actin cytoskeletons of thrombin-activated platelets. J Cell Biol 1993; 121: 1329-42.
- 15 Fischer TH, Gatling MN, Lacal JC, White II GC. Rap1b, a cAMP-dependent protein kinase substrate, associates with the platelet cytoskeleton. J Biol Chem 1990; 265: 19405-8.
- 16 Torti M, Ranaschi G, Sinigaglia F, Lapetina EG, Balduini C. Association of the low molecular weight GTP-binding protein rap2B with the cytoskeleton during platelet aggregation. Proc Nat’l Acad Sci 1993; 90: 7553-7.
- 17 Zhang J, King WG, Dillon S, Hall A, Feig L, Rittenhouse SE. Activation of platelet phosphatidylinositol 3-kinase requires the small GTP-binding protein Rho. J Biol Chem 1993; 268: 22251-8.
- 18 Ramaschi G, Sinigaglia F, Torti M, Balduini C. Effect of cAMP on the association of low molecular weight GTP-binding proteins with platelet cytoskeleton. Biochim Biophys Acta 1994; 1199: 20-6.
- 19 Ridley AJ, Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 1992; 70: 401-0.
- 20 Ridley AJ, Paterson HF, Johnston CL, Diekmann D, Hall A. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell 1992; 70: 469-72.
- 21 Fischer TH, Gatling MN, McCormick FA, Duffy CM, White II GC. Incorporation of rap1b into the platelet cytoskeleton is dependent on thrombin activation and extracellular calcium. J Biol Chem 1994; 269: 17257-61.
- 22 Fischer TH, White II GC. Partial purification and characterization of thrombolamban, a 22,000 dalton cAMP-dependent protein kinase substrate in platelets. Biochem Biophys Res Commun 1987; 149: 700-6.
- 23 Fitzgerald LA, Leung B, Phillips DR. A method for purifying the platelet membrane glycoprotein GPIIb/IIIa complex. Anal. Biochem. 1985; 151: 169-77.
- 24 Lacal J, Aaronson S. Ras p21 deletion mutants and monoclonal antibodies as tools for localization of regions of relevant p21 function. Proc Nat’l Acad Sci 1986; 83: 5400-4.
- 25 White II GC, Workman Jr EF, Lundblad RL. Thrombin binding to thrombasthenic platelets. J Lab Clin Med 1978; 91: 76-82.
- 26 White II G.C., Barton D.W., White T.E.. and Fischer, T.H. Cyclic AMP-dependent protein kinase does not increase calcium transport in platelet microsomes. Thrombos. Res. 1989; 56: 575-81.
- 27 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-3.
- 28 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Nat’l. Acad. Sci. 1979; 76: 4350-4.
- 29 Torti M, Ramaschi G, Sinigaglia F, Lapetina EG, Balduini C. Glycoprotein IIb-IIIa and the translocation of rap2B to the platelet cytoskeleton. Proc Nat’l Acad Sci 1994; 91: 4239-43.
- 30 Shattil SJ, Hoxie JA, Cunningham M, Brass LF. Changes in the platelet membrane glycoprotein IIb-IIIa complex during activation. J Biol Chem 1985; 260: 11107-14.