Thromb Haemost 1994; 72(05): 762-769
DOI: 10.1055/s-0038-1648955
Original Article
Schattauer GmbH Stuttgart

Expression of Platelet Membrane Glycoprotein V in Human Megakaryocytes and Megakaryocytic Cell Lines: A Study Using a Novel Monoclonal Antibody against GPV

Toshiro Takafuta
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Kingo Fujirmura
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Hironori Kawano
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Masaaki Noda
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Tetsuro Fujimoto
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Kenji Oda
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Takeshi Shimomura
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
,
Atsushi Kuramoto
The Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima, Japan
› Author Affiliations
Further Information

Publication History

Received 28 March 1994

Accepted after revision 07 July 1994

Publication Date:
06 July 2018 (online)

Summary

Glycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.

 
  • References

  • 1 Mosher DF, Vaheri A, Choate JJ, Gahmberg CG. Action of thrombin on surface glycoproteins of human platelets. Blood 1979; 53: 437-445
  • 2 Phillips DR, Agin PP. Platelet plasma membrane glycoproteins. Identification of a proteolytic substrate for thrombin. Biochem Biophys Res Commun 1977; 75: 940-947
  • 3 Bemdt MC, Phillips DR. Purification and preliminary physiochemical characterization of human platelet membrane glycoprotein V. J Biol Chem 1981; 256: 59-65
  • 4 Bienz D, Schnippering W, Clemetson KJ. Glycoprotein V is not the thrombin activation receptor on human blood platelets. Blood 1986; 68: 720-725
  • 5 Vu T-K, Hung D, Wheaton V, Coughlin S. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-1068
  • 6 Shimomura T, Fujimura K, Maehama S, Takemoto M, Oda K, Fujimoto T, Oyama R, Suzuki M, Ichihara-Tanaka K, Titani K, Kuramoto A. Rapid purification and characterization of human platelet glycoprotein V. Blood 1990; 75: 2349-2356
  • 7 Roth GJ, Church TA, McMullen BA, Williams ST. Human platelet glycoprotein V: A surface leucine-rich glycoprotein related to adhesion. Biochem Biophys Res Commun 1990; 170: 153-161
  • 8 Modderman PW, Admiraal LG, Sonnenberg A, von dem Borne AEGKr. Glycoprotein V and Ib-IX form a non covalent complex in the platelet membrane. J Biol Chem 1992; 267: 364-369
  • 9 Roth GJ. Development relationships: arterial platelet adhesion, glycoprotein Ib, and leucine-rich glycoproteins. Blood 1991; 77: 05-19
  • 10 Hickey MJ, Hagen FS, Yagi M, Roth GJ. Human platelet glycoprotein V: characterization of the polypeptide and the related Ib-V-IX receptor system of adhesive, leucine-rich glycoproteins. Proc Natl Acad Sci USA 1993; 90: 8327-8331
  • 11 Jamieson GA, Okumura T. Reduced thrombin binding and aggregation in Bemard-Soulier syndrome platelets. J Clin Invest 1978; 61: 861-864
  • 12 Nurden AT, Dupuis D, Kunicki TJ, Caen JP. Analysis of the glycoprotein and protein composition of Bemard-Soulier platelets by single and two-dimensional sodium dodecyl sulfate-polyaerylamide gel electrophoresis. J Clin Invest 1981; 67: 1431-1440
  • 13 Clemetson KJ, McGregor JL, James E, Dechavanne M, Luscher EF. Characterization of the platelet membrane glycoprotein abnormalities in Bernard-Soulier syndrome and comparison with normal by surface-labeling techniques and high-resolution two-dimensional gel electrophoresis. J Clin Invest 1982; 70: 304-311
  • 14 Berndt MC, Gregory C, Chong BH, Zola H, Castaldi PA. Additional glycoprotein defects in Bemard-Soulier’s syndrome: Confirmation of genetic basis by parental analysis. Blood 1983; 62: 800-807
  • 15 George JN, Nurden AT, Phillips DR. Molecular defects in interactions of platelets with the vessel wall. N Engl J Med 1984; 311: 1084-1098
  • 16 Debili N, Kieffer N, Nakazawa M, Guichard J, Titeux M, Cramer E, Breton-Gorius J, Vainchenker W. Expression of platelet glycoprotein Ib by cultured human megakaryocytes: Ultrastructural localization and biosynthesis. Blood 1990; 76: 368-376
  • 17 Breton-Gorius J, Vainchenker W. Expression of platelet proteins during the in vitro and in vivo differentiation of megakaryocytes and morphological aspects of their maturation. Semin in Hematol 1986; 23: 43-67
  • 18 Kieffer N, Debili N, Wicki A, Titeux M, Henri A, Mishal Z, Breton-Gorius J, Vainchenker W, Clementson KJ. Expression of platelet glycoprotein Ibα in HEL cells. J Biol Chem 1986; 261: 15854-15862
  • 19 Takeuchi S, Sugito S, Uemura Y, Miyagi T, Kubonishi I, Taguchi H, Enzan H, Ohtsuki Y, Miyoshi I. Acute megakaryoblastic leukemia: Establishment of a new cell line (MKPL-1) in vitro and in vivo. Leukemia 1992; 6: 588-594
  • 20 Lanza F, Morale M, La Salle C, Cazenave JP, Clemetson KJ, Shimomura T, Philips DR. Cloning and characterization of the gene encoding the human platelet glycoprotein V. J Biol Chem 1993; 268: 20801-20807
  • 21 Fujimura K, Takafuta T, Kawano H, Shimomura T, Fujimoto T, Oda K, Titani K, Kuramoto A. Analysis of GPV by using a novel monoclonal anti GPV antibody. Thromb Haemost 1993; 69: 1335 (abstr)
  • 22 Komatsu N, Nakauchi H, Miwa A, Ishihara T, Eguchi M, Moroi M, Okada M, Sato Y, Wada H, Yawata Y, Suda T, Miura Y. Establishment and characterization of a human leukemic cell line with megakaryocytic features. Cancer Res 1991; 51: 341-348
  • 23 Ishibashi T, Ruggeri ZM, Harker LA, Burstein SA. Separation of human megakaryocytes by state of differentiation on continuous gradients of Percoll. Blood 1986; 67: 1286-1292
  • 24 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-159
  • 25 George JN, Pickett EB, Saucerman S, McEver RP, Kunicki TJ, Kieffer N, Newman PJ. Platelet surface glycoproteins. Studies on resting and activated platelets and platelet membrane microparticles in normal subjects, and observations in patients during adult respiratory distress syndrome and cardiac surgery. J Clin Invest 1986; 78: 340-348
  • 26 Yam LT, Li CY, Crosby WH. Cytochemical identification of monocytes and granulocytes. Am J Clin Pathol 1971; 55: 283-290
  • 27 Hermine O, Mayeux P, Titeux M, Mitjavila MT, Casadevall N, Guichard J, Komatsu N, Suda T, Miura Y, Vainchenker W, Breton-Gorius J. Granulocyte-macrophage colony-stimulating factor and erythropoietin act competitively to induce two different programs of differentiation in the human pluripotent cell line. Blood 1992; 80: 3060-3069