Biopolym. Cell. 2015; 31(2):104-108.
Structure and Function of Biopolymers
Lys-plasminogen stimulates vitronectin exposure on the platelet surface
1Zhernossekov D. D., 1Roka-Moiia Y. M., 1Tykhomyrov A. A., 1Grinenko T. V.
  1. Palladin Institute of Biochemistry, NAS of Ukraine
    9, Leontovycha Str., Kyiv, Ukraine, 01601

Abstract

Aim. To study the vitronectin exposure on the platelet surface in the presence of two forms of the plasminogen molecule: Lys- and Glu-plasminogens. Methods. Research was conducted on human platelets. Aggregometry was used to check the platelet vitality and cell response towards thrombin. To evaluate the influence of different plasminogen forms on the platelet vitronectin exposure we used the method of flow cytometry. Results. It was found that incubation of resting platelets with Lys-plasminogen increased the amount of vitronectin-positive cells, but did not affect significantly their fluorescent intensity. Thrombin stimulation led to an increase in both: the number of vitronectin-positive platelets and the signal of fluorescence at least by two times. The Lys-plasminogen adding to the suspension of washed platelets followed by the thrombin stimulation enhanced the vitronectin exposure on the platelet surface and increased the amount of vitronectin-positive cells as compared to the isolated thrombin stimulation. Glu-plasminogen had no effect on the vitronectin exposure in case of resting or stimulated platelets. Conclusions. Lys-plasminogen but not its Glu-form enhances the exposure of vitronectin on the platelet surface. We suggest that the binding of Lys-plasminogen to the surface platelet receptors may generate plasmin that leads to the activation of intracellular signaling cascade.
Keywords: plasminogen, platelets, platelet secretion, vitronectin, flow cytometry.

References

[1] Broos K, Feys HB, De Meyer SF, Vanhoorelbeke K, Deckmyn H. Platelets at work in primary hemostasis. Blood Rev. 2011;25(4):155-67.
[2] Ni H, Denis CV, Subbarao S, Degen JL, Sato TN, Hynes RO, Wagner DD. Persistence of platelet thrombus formation in arterioles of mice lacking both von Willebrand factor and fibrinogen. J Clin Invest. 2000;106(3):385-92.
[3] Reheman A, Gross P, Yang H, Chen P, Allen D, Leytin V, Freedman J, Ni H. Vitronectin stabilizes thrombi and vessel occlusion but plays a dual role in platelet aggregation. J Thromb Haemost. 2005;3(5):875-83.
[4] Stockmann A, Hess S, Declerck P, Timpl R, Preissner KT. Multimeric vitronectin. Identification and characterization of conformation-dependent self-association of the adhesive protein. J Biol Chem. 1993;268(30):22874-82.
[5] Seiffert D, Schleef RR. Two functionally distinct pools of vitronectin (Vn) in the blood circulation: identification of a heparin-binding competent population of Vn within platelet alpha-granules. Blood. 1996;88(2):552-60.
[6] Asch E, Podack E. Vitronectin binds to activated human platelets and plays a role in platelet aggregation. J Clin Invest. 1990;85(5):1372-8.
[7] Zhou A, Huntington JA, Pannu NS, Carrell RW, Read RJ. How vitronectin binds PAI-1 to modulate fibrinolysis and cell migration. Nat Struct Biol. 2003;10(7):541-4.
[8] Kost C, St?ber W, Ehrlich HJ, Pannekoek H, Preissner KT. Mapping of binding sites for heparin, plasminogen activator inhibitor-1, and plasminogen to vitronectin's heparin-binding region reveals a novel vitronectin-dependent feedback mechanism for the control of plasmin formation. J Biol Chem. 1992;267(17):12098-105.
[9] Roka-Moya YM, Zhernossekov DD, Grinenko TV. Plasminogen. plasmin influence on platelet aggregation. Biopolym Cell. 2012;28(5):352–6.
[10] Deutsch DG, Mertz ET. Plasminogen: purification from human plasma by affinity chromatography. Science. 1970;170(3962):1095-6.
[11] Leytin V, Mody M, Semple JW, Garvey B, Freedman J. Quantification of platelet activation status by analyzing P-selectin expression. Biochem Biophys Res Commun. 2000;273(2):565-70.
[12] Miles LA, Dahlberg CM, Plow EF. The cell-binding domains of plasminogen and their function in plasma. J Biol Chem. 1988;263(24):11928-34.
[13] Parker CJ, Stone OL, White VF, Bernshaw NJ. Vitronectin (S protein) is associated with platelets. Br J Haematol. 1989;71(2):245-52.
[14] Godier A, Hunt BJ. Plasminogen receptors and their role in the pathogenesis of inflammatory, autoimmune and malignant disease. J Thromb Haemost. 2013;11(1):26-34.