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Thromb Haemost 1996; 75(04): 623-629
DOI: 10.1055/s-0038-1650333
Original Article
Schattauer GmbH Stuttgart

The Lysosomal Granule Membrane Protein, Lamp-2, Is also Present in Platelet Dense Granule Membranes

S J Israels
The Departments of Pediatrics and Oral Biology and the Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada
,
E M McMillan
The Departments of Pediatrics and Oral Biology and the Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada
,
C Robertson
The Departments of Pediatrics and Oral Biology and the Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada
,
S Singhroy
The Departments of Pediatrics and Oral Biology and the Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada
,
A McNicol
The Departments of Pediatrics and Oral Biology and the Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada
› Author Affiliations
Further Information

Publication History

Received 31 May 1995

Accepted after revision 12 January 1996

Publication Date:
10 July 2018 (online)

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Summary

Lysosomal Associated Membrane Protein-2 (LAMP-2) is an inherent component of lysosomal granule membranes in diverse cell types, including platelets. We examined platelets for evidence of LAMP-2 in dense granule membranes as CD63 has previously been shown to be present in both lysosomal and dense granule membranes. Immunological techniques were used to examine the localization of LAMP-2 in control platelets and those from an individual with Hermansky-Pudlak syndrome (HPS), a condition characterised by platelet dense granule deficiency. Immunoblotting studies demonstrated that LAMP-2 was enriched in a dense granule preparation. Flow cytometry of thrombin-stimulated control platelets was consistent with biphasic surface expression of LAMP-2. The early expression was accompanied by dense granule, but minimal lysosomal granule, release. The late expression was accompanied by additional lysosomal granule release only. Thrombin stimulation of HPS platelets showed only late, lysosome-associated LAMP-2 expression. Immunoelectron microscopy indicated the presence of LAMP-2 in the membranes of serotonin-containing granules as identified by an anti-serotonin polyclonal antibody. These data indicate that LAMP-2 is present in the membranes of platelet dense granules in addition to lysosomal granules, and has a similar distribution to CD63.

 

  • References

  • 1 Gerrard JM, McNicol A, Saxena SP. Protein kinase C and granule membrane fusion in platelets. In: Signal Transduction during Biomembrane Fusion O’Day DH. (ed) San Diego: Academic Press; 1993: 121-145
  • 2 McNicol A, Israels SJ, Gerrard JM. Platelets. Recent Advances in Blood Coagulation Poller L. (ed) Edinburgh: Churchill Livingstone: 1993: 17-49
  • 3 Wencel-Drake JD, Plow EF, Kunicki TJ, Woods VL, Keller DM, Ginsberg MH. Localization of internal pools of membrane glycoproteins involved in platelet adhesive responses. Am J Pathol 1986; 124: 324-334
    PubMedSearch in Google Scholar
  • 4 Stenberg PE, McEver RP, Shuman MA, Jacques YV, Bainton DF. A platelet a-granule membrane protein (GMP-140) is expressed on the plasma membrane after activation. J Cell Biol 1985; 101: 880-886
    CrossrefPubMedSearch in Google Scholar
  • 5 Israels SJ, Gerrard JM, Jaques YV, McNicol A, Cham B, Nishibori M, Bainton DF. Platelet dense granule membranes contain both granulophysin and P-selectin (GMP-140). Blood 1992; 80: 143-152
    CrossrefPubMedSearch in Google Scholar
  • 6 Plow EF, Ginsberg MH. Cellular adhesion: GPIIb-IIIa as a prototypic adhesion receptor. In: Progress in Hemostasis and Thrombosis Coller BS. (ed) Philadelphia: WB Saunders; Vol. 9 1989: 117-156
  • 7 Hamburger SA, McEver RP. GMP-140 mediates adhesion of stimulated platelets to neutrophils. Blood 1990; 75: 550-554
    PubMedSearch in Google Scholar
  • 8 White JG. The dense bodies of human platelets: Inherent electron opacity of the serotonin storage particles. Blood 1969; 33: 598-606
    PubMedSearch in Google Scholar
  • 9 Israels SJ, McNicol A, Robertson C, Gerrard JM. Platelet storage pool deficiency: diagnosis in patients with prolonged bleeding times and normal platelet aggregation. Br J Haematol 1990; 75: 118-121
    CrossrefPubMedSearch in Google Scholar
  • 10 Nieuwenhuis HK, Akkerman JN, Sixma JJ. Patients with a prolonged bleeding time and normal aggregation tests may have storage pool deficiency: Studies on one hundred and six patients. Blood 1987; 70: 620-623
    CrossrefPubMedSearch in Google Scholar
  • 11 White JG. Platelet granule disorders. CRC Crit Rev Oncol Hematol 1986; 4: 337-377
    CrossrefPubMedSearch in Google Scholar
  • 12 Weiss HJ, Tschopp TB, Rogers J, Brand H. Studies of platelet 5-hydroxytryptamine (serotonin) in storage pool disease and albinism. J Clin Invest 1974; 54: 421-432
    CrossrefPubMedSearch in Google Scholar
  • 13 Weiss HJ, Lages B, Vicic W, Tsung LY, White JG. Heterogenous abnormalities of platelet dense granule ultrastructure in 20 patients with congenital storage pool deficiency. Br J Haematol 1993; 83: 282-295
    CrossrefPubMedSearch in Google Scholar
  • 14 Lloyd JV. Hereditary disorders of platelet function. Platelets 1993; 4: 243-251
    CrossrefPubMedSearch in Google Scholar
  • 15 Carty SE, Johnson RG, Scarpa A. Serotonin transport in isolated platelet granules. J Biol Chem 1981; 256: 11244-11250
    PubMedSearch in Google Scholar
  • 16 Rendu F, Lebret M, Danielian S, Fagard R, Levy-Toledano S, Fischer S. High pp60c’src level in human platelet dense bodies. Blood 1989; 73: 1545-1551
    PubMedSearch in Google Scholar
  • 17 Rendu F, Martin CramerE, Tenza D, Fischer S. Redistribution of granulophysin and src protein in normal and Gray platelets after activation. Platelets 1993; 4: 167-174
    CrossrefPubMedSearch in Google Scholar
  • 18 Ferrell JE, Noble JA, Martin GS, Jaques YV, Bainton DF. Intracellular localization of pp60c’src in human platelets. Oncogene 1990; 5: 1033-1036
    PubMedSearch in Google Scholar
  • 19 Sorisky A, Lages B, Weiss HJ, Rittenhouse SE. Human platelets deficient in dense granules contain normal amounts of pp60c src. Thromb Res 1992; 65: 77-83
    CrossrefPubMedSearch in Google Scholar
  • 20 Nieuwenhuis HK, van Oosterhout JJG, Rozenmuller E, van Iwaarden F, Sixma JJ. Studies with a monoclonal antibody against activated platelets: Evidence that a secreted 53,000 molecular weight lysosome-like granule protein is exposed on the surface of activated platelets in the circulation. Blood 1987; 70: 838-845
    CrossrefPubMedSearch in Google Scholar
  • 21 Metzelaar MJ, Wingaard LJ, Peters PJ, Sixma JJ, Nieuwenhuis HK, Clevers HC. CD63 antigen: A novel lysosomal membrane glycoprotein cloned by a screening procedure for intracellular antigens in eukaryotic cells. J Biol Chem 1991; 266: 3239-3245
    CrossrefPubMedSearch in Google Scholar
  • 22 Demetrick DJ, Herlyn D, Tretiak M, Creasey D, Cleves H, Donoso LA, Vennegoor CJGM, Dixon WT, Jerry LM. ME491 melanoma-associated glycoprotein family: Antigenic identity of ME491, NKI/C-3, neuroglandu-lar antigen (NGA), and CD63 proteins. J Natl Cancer Inst 1992; 84: 422-429
    CrossrefPubMedSearch in Google Scholar
  • 23 Metzelaar MJ, Schuurman HJ, Heijnen HFG, Sixma JJ, Nieuwenhuis HK. Biochemical and immunohistochemical characteristics of CD62 and CD63 monoclonal antibodies. Virchows Archiv B Cell Pathol 1991; 61: 269-277
    PubMedSearch in Google Scholar
  • 24 Hamamoto K, Ohga S, Nomura S, Yasunaga K. Cellular distribution of CD63 antigen in platelets and in three megakaryocytic cell lines. Histochem J 1994; 26: 367-375
    CrossrefPubMedSearch in Google Scholar
  • 25 Hotta H, Ross AH, Huebner K, Isobe M, Wendebom S, Chao MV, Ricciar-di RP, Tsujimoto Y, Croce CM, Koprowski H. Molecular cloning and characterization of an antigen associated with early stages of melanoma tumor progression. Cancer Res 1988; 48: 2955-2962
    PubMedSearch in Google Scholar
  • 26 Vischer UM, Wagner DD. CD63 is a component of Weibel-Palade bodies of human endothelial cells. Blood 1993; 82: 1184-1191
    PubMedSearch in Google Scholar
  • 27 Nishibori M, Cham B, McNicol A, Shalev A, Jain N, Gerrard JM. The protein CD63 in platelet dense granules, is deficient in a patient with Her-mansky-Pudlak syndrome, and appears identical to granulophysin. J Clin Invest 1993; 91: 1775-1782
    CrossrefPubMedSearch in Google Scholar
  • 28 Gerrard JM, Lint D, Sims PJ, Weidmer T, Fugate RD, McMillan E, Robertson C, Israels SJ. Identification of a platelet dense granule membrane protein that is deficient in a patient with the Hermansky-Pudlak syndrome. Blood 1991; 77: 101-112
    PubMedSearch in Google Scholar
  • 29 Febbraio M, Silverstein RL. Identification and characterization of LAMP-1 as an activation dependent platelet surface glycoprotein. J Biol Chem 1990; 265: 18531-18537
    CrossrefPubMedSearch in Google Scholar
  • 30 Silverstein RL, Febbraio M. Identification of lysosome-associated membrane protein-2 as an activation-dependent platelet surface glycoprotein. Blood 1992; 80: 1470-1475
    CrossrefPubMedSearch in Google Scholar
  • 31 Metzelaar MJ, Clevers HC. Lysosomal membrane glycoproteins in platelets. Thromb Haemost 1992; 68: 378-382
    Thieme ConnectPubMedSearch in Google Scholar
  • 32 Fukuda M. Lysosomal membrane glycoproteins: Structure, biosynthesis, and intracellular trafficking. J Biol Chem 1991; 266: 21327-21330
    PubMedSearch in Google Scholar
  • 33 Murayama T, Kajiyama Y, Nomura Y. Histamine-stimulated and GTP-binding protein-mediated phospholipase A2 activation in rabbit platelets. J Biol Chem 1990; 265: 4290-4295
    PubMedSearch in Google Scholar
  • 34 Gerrard JM, Beattie LL, Park J, Israels SJ, McNicol A, Lint DW, Cragoe EJ. A role for protein kinase C in the membrane fusion necessary for platelet granule secretion. Blood 1989; 74: 2405-2413
    PubMedSearch in Google Scholar
  • 35 Hoehn SK, Kanfer JN. L-ascorbic acid and lysosomal acid hydrolase activities of guinea pig liver and brain. Can J Biochem 1978; 56: 352-356
    CrossrefPubMedSearch in Google Scholar
  • 36 Rendu F, Lebret M, Nurden AT, Caen JP. Initial characterization of human platelet mepacrine-labeled granules isolated using a short metrizamide gradient. Br J Haematol 1982; 52: 241-251
    CrossrefPubMedSearch in Google Scholar
  • 37 Tokayasu KT. Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem J 1989; 21: 163-171
    CrossrefPubMedSearch in Google Scholar
  • 38 Stenberg PE, Shuman MA, Levine SP, Bainton DF. Optimal techniques for immunocytochemical demonstration of fl-thromboglobulin, platelet factor 4, and fibrinogen in the alpha granules of unstimulated platelets. Histochem J 1984; 16: 983-1001
    CrossrefPubMedSearch in Google Scholar
  • 39 Steinbusch HWM, Verhofstad AAJ, Joosten HWJ. Localization of serotonin in the central nervous system by immunohistochemistry: Description of a specific and sensitive technique and some applications. Neurosci 1978; 3: 811-819
    CrossrefPubMedSearch in Google Scholar
  • 40 Gaudin-Chazel G, Portalier P, Barrit MC, Puizillout JJ. Serotonin-like immunoreactivity in paraffin sections of the nodose ganglia of the cat. Neurosci Lettr 1982; 33: 169-172
    PubMedSearch in Google Scholar
  • 41 McNicol A, Israels SJ, Robertson C, Gerrard JM. The empty sack syndrome: A platelet storage pool deficiency associated with empty dense granules. Br J Haematol 1994; 86: 574-582
    CrossrefPubMedSearch in Google Scholar
  • 42 Gerrard JM, McNicol A, Klassen D, Israels SJ. Protein phosphorylation and its relation to platelet function. In: Blood Cells and Arteries in Hypertension and Atherosclerosis Meyer P, Marche P. (eds) New York: Raven Press; 1989: 93-114
  • 43 Williams MA, Fukuda M. Accumulation of membrane glycoproteins in lysosomes requires a tyrosine residue at a particular position in the cytoplasmic tail. J Cell Biol 1990; 111: 955-966
    CrossrefPubMedSearch in Google Scholar
  • 44 Hieber AD, Christie DL. Characterization of glycoprotein II from bovine adrenal medullary chromaffin granules. J Biol Chem 1993; 268: 11073-11078
    PubMedSearch in Google Scholar
  • 45 Peters PJ, Borst J, Oorschot V, Fukuda M, Krahenbuhl O, Tschopp J, Slot JW, Geuze HJ. Cytotoxic T lymphocyte granules are secretory lysosomes, containing both perforin and granzymes. J Exp Med 1991; 173: 1099-1109
    CrossrefPubMedSearch in Google Scholar
  • 46 Kornfeld S, Mellman I. The biogenesis of lysosomes. Annu Rev Cell Biol 1989; 5: 483-525
    CrossrefPubMedSearch in Google Scholar