Fibrinogen Binding Is Independent of an Increase in Intracellular Calcium Concentration in Thrombin Degranulated Platelets

Fabio M Pulcinelli; James L Daniel; Silvia Riondino; Pier Paolo Gazzaniga; Leon Salganicoff

doi:10.1055/s-0038-1653769

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1995; 73(02): 304-308
DOI: 10.1055/s-0038-1653769

Original Article

Platelets

Schattauer GmbH Stuttgart

Fibrinogen Binding Is Independent of an Increase in Intracellular Calcium Concentration in Thrombin Degranulated Platelets

Fabio M Pulcinelli

The Dept. of Experimental Medicine, University La Sapienza, Rome, Italy

,

James L Daniel

¹The Dept. of Pharmacology, School of Medicine, Temple University, Philadelphia, PA, USA

,

Silvia Riondino

The Dept. of Experimental Medicine, University La Sapienza, Rome, Italy

,

Pier Paolo Gazzaniga

The Dept. of Experimental Medicine, University La Sapienza, Rome, Italy

,

Leon Salganicoff

¹The Dept. of Pharmacology, School of Medicine, Temple University, Philadelphia, PA, USA

› Author Affiliations

Abstract

PDF Download

Summary

In a suspension of thrombin degranulated platelets (TDP), ADP and epinephrine can induce platelet aggregation, whereas the synthetic agonist of the thromboxane/endoperoxide receptor U46619 causes only shape change. However, U46619 can enhance platelet aggregation induced by ADP and epinephrine. In this paper, we have measured fibrinogen binding in relation to phospholipase C(PLC) activation and calcium mobilization in TDP activated by ADP, epinephrine and U46619.

ADP caused fibrinogen binding in TDP but neither activated PLC nor caused a calcium mobilization. The requirement for ADP in inducing exposure of fibrinogen binding sites was not absolute since the combination of epinephrine and U46619 produced an increase in fibrinogen binding. U46619 caused significant PLC activation and cytosolic calcium release but not fibrinogen binding. These results suggest that in TDP the exposure of fibrinogen binding sites, after agonist activation, is independent of both PLC activation and calcium mobilization.

PDF (1111 kb)

References

References
1 Pytela R, Pierschbacher MD, Ginsberg MH, Plow EF, Ruoslahti E. Platelet membrane glycoprotein IIb/IIIa: member of a family of Arg-Gly-Asp-specific adhesion receptors. Sci 1986; 231: 1559-1562
2 Gartner TK, Bennet JS. The tetrapeptide analogue of the cell attachment site of fibronectin inhibits platelet aggregation and fibrinogen binding to activated platelets. J Biol Chem 1985; 260: 11891-11894
3 Ginsberg MH, Loftus JC, Plow EF. Cytoadhesins, integrins, and platelets. Thromb Haemost 1988; 59: 1-6
4 Shattil SJ. Regulation of platelet anchorage and signaling by integrin αIIb-β3. Thromb Haemost 1993; 70: 224-228
5 Morinelli TA, Niewiarowski S, Kornecki E, Figures WR, Wachtfogel Y, Colman RW. Platelet aggregation and exposure of fibrinogen receptors by prostaglandin endoperoxide analogues. Blood 1983; 61: 41-49
6 Pulcinelli FM, Daniel JL, Riondino S, Gazzaniga PP, Russo PP, Salganicoff L. A method to prepare thrombin degranulated platelets. Platelets 1993; 4: 212-218
7 Huang EM, Detwiler TC. Stimulus-response coupling mechanisms. In: Biochemistry of platelets. Phillips DR, Shuman MA. eds. New York: Academic Press; 1986. pp 1-68
8 Ashby B, Daniel JL, Smith JB. Mechanisms of platelet activation and inhibition. In: Hematology/Oncology Clinics of North America. Platelets in health and disease Colman RW, Rao AK. eds. Philadelphia: Saunders Company; 1990. pp 1-26
9 Aster RH, Jandel JH. Platelet sequestration in man. I. Methods J Clin Invest 1964; 43: 834-855
10 Marguerie GA, Plow EF, Edgington TS. Human platelets possess an inducible and saturable receptor specific for fibrinogen. J Biol Chem 1979; 254: 5357-5363
11 Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca²⁺ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440-3445
12 Pulcinelli FM, Gazzaniga PP, Salganicoff L. Use of Zn-pyrophosphatase in the high-performance liquid chromatographic analysis of cell extracts containing ³²P-labelled inositol phosphates. J Chromatogr 1992; 575: 51-55
13 Daniel JL, Dangelmaier CA, Smith JB. Formation and metabolism of inositol 1, 4, 5-triphosphate in human platelets. Biochem J 1987; 246: 109-114
14 Haslam RJ. Role of the adenosine diphosphate in the aggregation of human blood platelets by thrombin. Nature 1964; 202: 765
15 Ingerman CM, Smith JB, Shapiro S, Sedar A, Silver MJ. Hereditary abnormality of platelet aggregation attributable to nucleotide storage pool deficiency. Blood 1978; 52: 332-344
16 MacFarlane DE, Mills DC B. The effect of ATP: evidence against the central role of released ADP in primary aggregation. Blood 1975; 46: 309-320
17 Shattil SJ, Budzynski A, Scrutton MC. Epinephrine induces platelet fibrinogen receptor expression, fibrinogen binding, and aggregation in whole blood in the absence of other excitatory agonists. Blood 1989; 73: 150-158
18 Daniel JL, Dangelmaier CA, Selak M, Smith JB. ADP stimulates IP3 formation in human platelets. FEBS Lett 1986; 206: 299-303
19 Vickers JD, Kinlough-Rathbone RL, Packham MA, Mustard JF. Inositol phospholipid metabolism in human platelets stimulated by ADP. Eur J Biochem 1990; 193: 521-528
20 Ginsberg MH, Xiaoping D, O'Toole TE, Loftus JC, Plow EF. Platelet- integrins. Thromb Haemost 1993; 70: 87-93
21 Parise LV, Criss AB, Nannizzi L, Wardell MR. Glycoprotein IIIa is phosphorylated in intact human platelets. Blood 1990; 75: 2363-2368
22 Hillery CA, Smyth SS, Parise LV. Phosphorylation of human platelet glycoprotein IIIa (GPIIIa). Dissociation from fibrinogen binding receptor activation and phosphorylation of GPIIIa in vitro J Biol Chem 1991; 266: 14663-14669
23 Shattil SJ, Cunningham M, Wiedmer T, Zhao J, Sims PJ, Brass LF. Regulation of glycoprotein IIb-IIIa receptor function studied with platelets permeabilized by the pore-forming complement proteins C5b-9. J Biol Chem 1992; 267: 18424-18431
24 Morii N, Teru-uchi T, Tominaga T, Kumagai N, Kozaki S, Ushikubi F, Narumiya S. A rho gene product in human blood platelets. II. Effect of the ADP-ribosylation by botulinum C3 ADP-ribosyltransferase on platelet aggregation J Biol Chem 1992; 267: 20921-20926