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Thromb Haemost 2001; 86(03): 887-893
DOI: 10.1055/s-0037-1616152
DOI: 10.1055/s-0037-1616152
Review Articles
The 33-kDa Platelet α-granule Membrane Protein (GMP-33) Is an N-terminal Proteolytic Fragment of Thrombospondin
This work was financially supported by the Dutch Heart Foundation (NHS) (grant no. 43.051).
Weitere Informationen
Publikationsverlauf
Received
12. Dezember 2000
Accepted after revision
07. Mai 2001
Publikationsdatum:
14. Dezember 2017 (online)
Summary
GMP-33 is a platelet membrane associated protein that is recognised by RUU-SP 1.77, an antibody raised against activated platelets. GMP-33 is predominantly associated with the membrane of platelet α-granules and it is translocated to the plasma membrane upon platelet activation (Metzelaar et al. Blood 1992; 79: 372-9). In this study we have isolated the protein by immunoaffinity chromatography. The N-terminus was sequenced and was identical to the N-terminal sequence of human thrombospondin. The protein was N-glycosylated and bound to heparin as would be expected of the N-terminal part of thrombospondin. RUU-SP 1.77 reacted only with reduced thrombospondin. Plasmin and trypsin digestion of thrombospondin yielded fragments of approximately the same size as GMP 33 that reacted with RUU-SP 1.77 after reduction. No evidence for alternative splicing was found. We postulate that GMP 33 is an N-terminal proteolytic fragment of thrombospondin that is membrane associated.
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References
- 1 Metzelaar MJ, Heijnen HF, Sixma JJ, Nieuwenhuis HK. Identification of a 33-Kd protein associated with the alpha-granule membrane (GMP-33) that is expressed on the surface of activated platelets. Blood 1992; 79: 372-9.
- 2 Lawler J, Hynes RO. The structure of human thrombospondin, an adhesive glycoprotein with multiple calcium-binding sites and homologies with several different proteins. J Cell Biol 1986; 103: 1635-48.
- 3 Gartner TK, Walz DA, Aiken M, Starr-Spires L, Ogilvie ML. Antibodies against a 23Kd heparin binding fragment of thrombospondin inhibit platelet aggregation. Biochem Biophys Res Commun 1984; 124: 290-5.
- 4 Metzelaar MJ, Korteweg J, Sixma JJ, Nieuwenhuis HK. Comparison of platelet membrane markers for the detection of platelet activation in vitro and during platelet storage and cardiopulmonary bypass surgery. J Lab Clin Med 1993; 121: 579-87.
- 5 Fauvel J, Chap H, Roques V, Levy-Toledano S, Douste-Blazy L. Biochemical characterization of plasma membranes and intracellular membranes isolated from human platelets using Percoll gradients. Biochim Biophys Acta 1986; 856: 155-64.
- 6 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970; 227: 680-5.
- 7 Edman P, Begg G. A protein sequenator. Eur J Biochem 1967; 1: 80-91.
- 8 Aiken ML, Ginsberg MH, Plow EF. Divalent cation-dependent and independent surface expression of thrombospondin on thrombin-stimulated human platelets. Blood 1987; 69: 58-64.
- 9 Morandi V, Edelman L, Legrand YJ, Legrand C. Characterization of a novel monoclonal antibody (V58A4) raised against a recombinant NH2-terminal heparin-binding fragment of human endothelial cell thrombospondin. FEBS Lett 1994; 346: 156-60.
- 10 Fugman DA, Witte DP, Jones CL, Aronow BJ, Lieberman MA. In vitro establishment and characterization of a human megakaryoblastic cell line. Blood 1990; 75: 1252-61.
- 11 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-9.
- 12 Sambrook J, Fritsch EF, Maniatis T. Molecular cloning, a laboratory manual. Cold Spring Harbor: Cold Spring Harbor Laboratory; 1989
- 13 Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-13.
- 14 Frohman MA, Dush MK, Martin GR. Rapid production of full-length cDNAs from rare transcripts: Amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 1988; 85: 8998-9002.
- 15 Schaefer BC. Revolutions in rapid amplification of cDNA ends: New strategies for polymerase chain reaction cloning of full-length cDNA ends. Anal Biochem 1995; 227: 255-73.
- 16 Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Bio. 1990; 215: 403-10.
- 17 Dixit VM, Grant GA, Santoro SA, Frazier WA. Isolation and characterization of a heparin-binding domain from the amino terminus of platelet thrombospondin. J Biol Chem 1984; 259: 10100-5.
- 18 Lawler J. The structure and functional properties of thrombospondin. Blood 1986; 67: 1197-209.
- 19 George JN, Lyons RM, Morgan RK. Membrane changes associated with platelet activation. Exposure of actin on the platelet surface after thrombin-induced secretion. J Clin Invest 1980; 66: 1-9.
- 20 Legrand C, Thibert V, Dubernard V, Begault B, Lawler J. Molecular requirements for the interaction of thrombospondin with thrombin-activated human platelets: modulation of platelet aggregation. Blood 1992; 79: 1995-2003.
- 21 Bornstein P, Sage EH. Thrombospondins. Methods Enzymol 1994; 245: 62-85.
- 22 Lawler J, Cohen AM, Chao FC, Moriarty DJ. Thrombospondin in essential thrombocythemia. Blood 1986; 67: 555-8.
- 23 Bornstein P. Diversity of function is inherent in matricellular proteins: An appraisal of thrombospondin 1. J Cell Biol 1995; 130: 503-6.
- 24 Reed MJ, Iruela-Arispe L, O’Brien ER, Truong T, LaBell T, Bornstein P, Sage EH. Expression of thrombospondins by endothelial cells – Injury is correlated with TSP-1. Am J Pathol 1995; 147: 1068-80.
- 25 Bornstein P, O’Rourke K, Wikstrom K, Wolf FW, Katz R, Li P, Dixit VM. A second, expressed thrombospondin gene (Thbs2) exists in the mouse genome. J Biol Chem 1991; 266: 12821-4.
- 26 Good DJ, Polverini PJ, Rastinejad F, Le Beau MM, Lemons RS, Frazier WA, Bouck NP. A tumor suppressor-dependent inhibitor of angiogenesis is immunologically and functionally indistinguishable from a fragment of thrombospondin. Proc Natl Acad Sci USA 1990; 87: 6624-8.
- 27 Rabhi Sabile S, Thibert V, Legrand C. Thrombospondin peptides inhibit the secretion-dependent phase of platelet aggregation. Blood Coag Fibrinolysis 1996; 7: 237-40.
- 28 Agbanyo FR, Sixma JJ, De Groot PG, Languino LR, Plow EF. Thrombospondin-platelet interactions. Role of divalent cations, wall shear rate, and platelet membrane glycoproteins. J Clin Invest 1993; 92: 288-96.
- 29 Gawaz M, Ott I, Reininger AJ, Heinzmann U, Neumann FJ. Agglutination of isolated platelet membranes. Arterioscler Thromb Vasc Biol 1996; 16: 621-7.
- 30 Murphy Ullrich JE, Gurusidappa S, Frazier WA, Hook M. Heparin binding peptides from thrombospondins 1 and 2 contain focal adhesion-labilizing activity. J Biol Chem 1993; 268: 26784-9.
- 31 Legrand C, Morandi V, Mendelovitz S, Shaked H, Hartman JR, Panet A. Selective inhibition of platelet macroaggregate formation by a recombinant heparin-binding domain of human thrombospondin. Arterioscler Thromb 1994; 14: 1784-91.