Localization and Translocation of MMP-2 during Aggregation of Human Platelets

Grzegorz Sawicki; Esmond J. Sanders; Eduardo Salas; Mieczyslaw Wozniak; Jose Rodrigo; Marek W. Radomski

doi:10.1055/s-0037-1615367

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1998; 80(05): 836-839
DOI: 10.1055/s-0037-1615367

Review Article

Schattauer GmbH

Localization and Translocation of MMP-2 during Aggregation of Human Platelets

Grzegorz Sawicki

¹From the Departments of Pharmacology, Edmonton, Canada

,

Esmond J. Sanders

²From the Departments of Physiology, University of Alberta, Edmonton, Canada

,

Eduardo Salas

³Division of Research, Lacer SA, Barcelona, Spain

,

Mieczyslaw Wozniak

⁴Department of Clinical Chemistry, Wroclaw University of Medicine, Wroclaw, Poland

,

Jose Rodrigo

⁵Department of Comparative Neuroanatomy, Cajal Institute, Madrid, Spain

,

Marek W. Radomski

¹From the Departments of Pharmacology, Edmonton, Canada

³Division of Research, Lacer SA, Barcelona, Spain

› Institutsangaben

Abstract

Summary

We have previously shown that human platelets express matrix metalloproteinase-2 (MMP-2) and that the release of this enzyme during platelet activation mediates the ADP- and thromboxane-independent part of aggregation. We have now used immunogold electron microscopy, flow cytometry, Western blot analysis and zymography methods to study the ultrastructural localization of MMP-2 in human washed platelets. Platelet aggregation was stimulated by collagen and the MMP-2 immunoreactivity of platelets was followed during various stages of aggregation. In resting platelets, MMP-2 was randomly distributed in the platelet cytosol without detectable association with platelet granules. Platelet aggregation caused the translocation of MMP-2 from the cytosol to the extracellular space. During the early stages of aggregation, MMP-2 remained in close association with the platelet plasma membrane. We conclude that the interactions of MMP-2 with platelet surface membranes mediate the aggregatory response induced by this enzyme.

Volltext

Referenzen

References
1 Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Cur Op Cell Biol 1995; 7: 728-35.
2 Ray JM, Stettler-Stevenson WG. The role of matrix metalloproteinases and their inhibitors in tumour invasion, metastasis and angiogenesis. Eur Resp J 1994; 7: 2062-72.
3 Sawicki G, Salas E, Murat J, Miszta-Lane H, Radomski MW. Release of gelatinase D during platelet activation mediates aggregation. Nature 1997; 386: 616-9.
4 Radomski MW, Moncada S. An improved method for washing of human platelets with prostacyclin. Thromb Res 1983; 30: 383-9.
5 Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mole Biol 1990; 215: 403-10.
6 Field GB. Solid phase peptide synthesis utilizing 9-fluorenylo-metoxy-carboxyl-amino acids. Int J Pept Prot Res 1990; 35: 161-214.
7 Harlow E, Lin D. Antibodies: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor; NY: 1988. pp. 82-3.
8 Jones D, Sawicki G, Wozniak M. Sequence, structure, and expression of a wasp venom protein with negatively charges signal peptide and a novel repeating internal structure. J Biol Chem 1992; 267: 14871-8.
9 Murphy G, Bretz U, Baggiolini M, Reynolds JJ. The latent collagenase and gelatinase of human polymorphonuclear neutrophil leucocytes. Biochem J 1980; 192: 517-25.
10 Dewald B, Bretz U, Baggiolini M. Release of gelatinase from a novel secre-tory compartment of human neutrophils. J Clin Invest 1982; 70: 518-25.
11 Kjeldsen L, Sengelov H, Lollike K, Nielsen MH, Borregaard N. Isolation and characterization of gelatinase granules from human neutrophils. Blood 1994; 83: 1640-9.
12 Borregaard N, Lollike K, Kjeldsen L, Sengelov H, Bastholm L, Nielsen MH, Bainton DF. Human neutrophil granules and secretory vesicles. Eur J Haematol 1993; 51: 187-98.
13 Kjeldsen L, Johnsen AH, Sengelov H, Borregaard N. Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. J Biol Chem 1993; 268: 10425-32.
14 Flower DR. The lipocalin protein family: a role in cell regulation. FEBS Lett 1994; 354: 7-11.
15 Poujol C, Nurden AT, Nurden P. Ultrastructural analysis of the distribution of the vitronectin receptor (α_vβ₃) in human platelets and megakaryocytes reveals an intracellular pool and labeling of the α-granule membrane. British J Haematol 1997; 97: 823-35.
16 Chanat E, Huttner WB. Milieu-induced, selective aggregation of regulated secretory proteins in the trans-Golgi network. J Cell Biol 1991; 115: 1505-19.
17 Siffert W, Akkerman JW. Activation of sodium-proton exchange is a prerequisite for Ca2+ mobilization in human platelets. Nature 1987; 325: 456-8.
18 Gullberg U, Andersson E, Garwicz D, Lindmark A, Olsson I. Biosynthesis, processing and sorting of neutrophil proteins: insight into neutrophil granule development. Eur J Haematol 1997; 58: 137-53.
19 White JG. Platelet membrane ultrastructure and its changes during platelet activation. In: Platelet Membrane Receptors: Molecular Biology, Immunology, Biochemistry and Pathology. Jamieson GA, ed. New York: Alan R, Liss, Inc.; 1988. pp. 1-32.
20 Clemetson KJ, Polgar J. Platelet adhesion and aggregation receptors. In: Platelets and their factors. Von Bruchhausen F, Walter U, eds. Berlin, Heidelberg, New York: Springer-Verlag; 1997. pp. 155-79.
21 Sato H, Okada Y, Seiki M. Membrane type matrix metalloproteinases (MT-MMPs) in cell invasion. Thromb Haemost 1997; 78: 497-500.
22 Vu T-KH, Hung DT, Wheaton VI, Coughlin SR. Molecular cloning of the functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-68.