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J Clin Invest. 1993 April; 91(4): 1775–1782.
PMCID: PMC288158

The protein CD63 is in platelet dense granules, is deficient in a patient with Hermansky-Pudlak syndrome, and appears identical to granulophysin.


The levels and expression of the proteins CD63 and granulophysin in platelets from control and from a Hermansky-Pudlak syndrome subject (a condition characterized by dense granule and lysosomal deficiencies and the accumulation of ceroid-like material in reticuloendothelial cells) were examined. Immunofluorescence studies indicated that anti-CD63 and anti-granulophysin antibodies recognized similar numbers of granules; coapplication of antibodies did not identify more granules than the individual antibodies. Significantly fewer granules were recognized in Hermansky-Pudlak syndrome platelets than in control using either antibody. Immunoblotting studies demonstrated that anti-CD63 and anti-granulophysin antibodies apparently recognize the same protein, which was deficient in Hermansky-Pudlak platelets. Analysis by fluorescence-activated cell sorter (FACS) showed biphasic expression of CD63 and granulophysin after thrombin stimulation of control but not Hermansky-Pudlak platelets. Anti-CD63 effectively blocked detection of the protein by anti-granulophysin using immunofluorescence, ELISA, immunoblotting, and FACS analysis. Amino-terminal sequencing over the first 37 amino acids revealed that granulophysin was homologous to CD63, melanoma antigen ME491, and pltgp40. These results suggest that granulophysin and CD63 are possibly identical proteins. This is the first report of a protein present in platelet dense granules, lysosomes, and melanocytes, but deficient in a patient with Hermansky-Pudlak syndrome.

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Selected References

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  • Gerrard JM. Platelet aggregation: cellular regulation and physiologic role. Hosp Pract (Off Ed) 1988 Jan 15;23(1):89–108. [PubMed]
  • Novak EK, Hui SW, Swank RT. The mouse pale ear pigment mutant as a possible animal model for human platelet storage pool deficiency. Blood. 1981 Jan;57(1):38–43. [PubMed]
  • Novak EK, Hui SW, Swank RT. Platelet storage pool deficiency in mouse pigment mutations associated with seven distinct genetic loci. Blood. 1984 Mar;63(3):536–544. [PubMed]
  • Novak EK, Swank RT. Lysosomal dysfunctions associated with mutations at mouse pigment genes. Genetics. 1979 May;92(1):189–204. [PubMed]
  • Brandt EJ, Elliott RW, Swank RT. Defective lysosomal enzyme secretion in kidneys of Chediak-Higashi (beige) mice. J Cell Biol. 1975 Dec;67(3):774–788. [PMC free article] [PubMed]
  • Reddington M, Novak EK, Hurley E, Medda C, McGarry MP, Swank RT. Immature dense granules in platelets from mice with platelet storage pool disease. Blood. 1987 May;69(5):1300–1306. [PubMed]
  • Swank RT, Reddington M, Howlett O, Novak EK. Platelet storage pool deficiency associated with inherited abnormalities of the inner ear in the mouse pigment mutants muted and mocha. Blood. 1991 Oct 15;78(8):2036–2044. [PubMed]
  • Nieuwenhuis HK, van Oosterhout JJ, Rozemuller 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 Sep;70(3):838–845. [PubMed]
  • Metzelaar MJ, Wijngaard PL, 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 Feb 15;266(5):3239–3245. [PubMed]
  • Atkinson B, Ernst CS, Ghrist BF, Herlyn M, Blaszczyk M, Ross AH, Herlyn D, Steplewski Z, Koprowski H. Identification of melanoma-associated antigens using fixed tissue screening of antibodies. Cancer Res. 1984 Jun;44(6):2577–2581. [PubMed]
  • Hotta H, Ross AH, Huebner K, Isobe M, Wendeborn S, Chao MV, Ricciardi 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 Jun 1;48(11):2955–2962. [PubMed]
  • Demetrick DJ, Herlyn D, Tretiak M, Creasey D, Clevers H, Donoso LA, Vennegoor CJ, Dixon WT, Jerry LM. ME491 melanoma-associated glycoprotein family: antigenic identity of ME491, NKI/C-3, neuroglandular antigen (NGA), and CD63 proteins. J Natl Cancer Inst. 1992 Mar 18;84(6):422–429. [PubMed]
  • Hardisty RM, Mills DC, Ketsa-Ard K. The platelet defect associated with albumism. Br J Haematol. 1972 Dec;23(6):679–692. [PubMed]
  • Witkop CJ., Jr Albinism. Adv Hum Genet. 1971;2:61–142. [PubMed]
  • Gerrard JM, Lint D, Sims PJ, Wiedmer 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 Jan 1;77(1):101–112. [PubMed]
  • Israels SJ, Gerrard JM, Jacques YV, McNicol A, Cham B, Nishibori M, Bainton DF. Platelet dense granule membranes contain both granulophysin and P-selectin (GMP-140). Blood. 1992 Jul 1;80(1):143–152. [PubMed]
  • Shalev A, Michaud G, Israels SJ, McNicol A, Singhroy S, McMillan EM, White JG, Witkop CJ, Nichols WL, Greenberg AH, et al. Quantification of a novel dense granule protein (granulophysin) in platelets of patients with dense granule storage pool deficiency. Blood. 1992 Sep 1;80(5):1231–1237. [PubMed]
  • Shalev A, Gerrard JM, Robertson C, Greenberg AH, Linial M. Sharing of antigenic epitopes between synaptophysin and granulophysin. J Cell Biochem. 1992 May;49(1):59–65. [PubMed]
  • Murayama T, Kajiyama Y, Nomura Y. Histamine-stimulated and GTP-binding proteins-mediated phospholipase A2 activation in rabbit platelets. J Biol Chem. 1990 Mar 15;265(8):4290–4295. [PubMed]
  • Kenney DM, Chao FC. Microtubule inhibitors alter the secretion of beta-glucuronidase by human blood platelets: involvement of microtubules in release reaction II. J Cell Physiol. 1978 Jul;96(1):43–52. [PubMed]
  • D'Souza SE, Ginsberg MH, Plow EF. Arginyl-glycyl-aspartic acid (RGD): a cell adhesion motif. Trends Biochem Sci. 1991 Jul;16(7):246–250. [PubMed]
  • Gerrard JM, Beattie LL, Park J, Israels SJ, McNicol A, Lint D, Cragoe EJ., Jr A role for protein kinase C in the membrane fusion necessary for platelet granule secretion. Blood. 1989 Nov 15;74(7):2405–2413. [PubMed]
  • Hoehn SK, Kanfer JN. L-Ascorbic acid and lysosomal acid hydrolase activities of guinea pig liver and brain. Can J Biochem. 1978 May;56(5):352–356. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Rendu F, Lebret M, Nurden AT, Caen JP. Initial characterization of human platelet mepacrine-labelled granules isolated using a short metrizamide gradient. Br J Haematol. 1982 Oct;52(2):241–251. [PubMed]
  • Weiss HJ, Chervenick PA, Zalusky R, Factor A. A familialdefect in platelet function associated with imapired release of adenosine diphosphate. N Engl J Med. 1969 Dec 4;281(23):1264–1270. [PubMed]
  • Rendu F, Maclouf J, Launay JM, Boinot C, Levy-Toledano S, Tanzer J, Caen J. Hermansky-Pudlak platelets: further studies on release reaction and protein phosphorylations. Am J Hematol. 1987 Jun;25(2):165–174. [PubMed]
  • Holmsen H, Day HJ. The selectivity of the thrombin-induced platelet release reaction: subcellular localization of released and retained constituents. J Lab Clin Med. 1970 May;75(5):840–855. [PubMed]
  • Holmsen H, Robkin L, Day HJ. Effects of antimycin A and 2-deoxyglucose on secretion in human platelets. Differential inhibition of the secretion of acid hydrolases and adenine nucleotides. Biochem J. 1979 Aug 15;182(2):413–419. [PubMed]
  • Azorsa DO, Hyman JA, Hildreth JE. CD63/Pltgp40: a platelet activation antigen identical to the stage-specific, melanoma-associated antigen ME491. Blood. 1991 Jul 15;78(2):280–284. [PubMed]
  • Pearson WR, Lipman DJ. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. [PubMed]
  • Abdelhaleem MM, Hatskelzon L, Dalal BI, Gerrard JM, Greenberg AH. Leukophysin: a 28-kDa granule membrane protein of leukocytes. J Immunol. 1991 Nov 1;147(9):3053–3059. [PubMed]

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