Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Molecular defects of the platelet P2 receptors

  • 122 Accesses

  • 17 Citations

Abstract

Human platelets express three types of P2 receptors, which play important roles in platelet function: P2X1, P2Y1 and P2Y12. Only patients with either quantitative or qualitative abnormalities of the platelet P2Y12 receptor have been well-characterized so far. Deficiencies of P2Y12 are associated with nucleotide deletions in the open-reading frame, frameshifts, and early truncation of the protein, or with a nucleotide substitution in the transduction initiation codon. Congenital dysfunctions of P2Y12 are associated with molecular defects involving the sixth trans-membrane domain or the adjacent third extracellular loop of the receptor, which identify a region of the protein whose integrity is necessary for normal receptor function. A mutation, predicting a lysine to glutamate (Lys174Glu) substitution was associated with decreased ligand binding to the receptor, suggesting that it is responsible for disruption of the adenosine diphosphate (ADP)-binding site of the receptor. Patients with P2Y12 defects display a mild-to-moderate bleeding diathesis, characterized by mucocutaneous bleedings and excessive post-surgical and post-traumatic blood loss. Defects of P2Y12 should be suspected when ADP, even at high concentrations (≥10 μM), is unable to induce full, irreversible platelet aggregation. Tests that evaluate the degree of inhibition of adenylyl cyclase by ADP should be used to confirm the diagnosis.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Cattaneo M, Gachet C (1999) ADP receptors and clinical bleeding disorders. Arterioscler Thromb Vasc Biol 19:2281–2285

  2. 2.

    Moore DJ, Murdock PR, Watson JM, Faull RL, Waldvogel HJ, Szekeres PG, Wilson S, Freeman KB, Emson PC (2003) GPR105, a novel Gi/o-coupled UDP-glucose receptor expressed on brain glia and peripheral immune cells, is regulated by immunologic challenge: possible role in neuroimmune function. Brain Res Mol Brain Res 118:10–23

  3. 3.

    von Kügelgen I (2006) Pharmacological profiles of cloned mammalian P2Y-receptor subtypes. Pharmacol Ther 110:415–432

  4. 4.

    Dovlatova N, Wijeyeratne YD, Fox SC, Manolopoulos P, Johnson AJ, White AE, Latif ML, Ralevic V, Heptinstall S (2008) Detection of P2Y(14) protein in platelets and investigation of the role of P2Y(14) in platelet function in comparison with the EP(3) receptor. Thromb Haemost 100:261–270

  5. 5.

    Huang EM, Detwiler TC (1986) In: Phillips DR, Shuman MA (eds) Biochemistry of platelets. Academic, Orlando, p 68

  6. 6.

    Falcon CR, Cattaneo M, Ghidoni A, Mannucci PM (1993) The in vitro production of thromboxane B2 by platelets of diabetic patients is normal at physiological concentrations of ionised calcium. Thromb Haemost 70:389–392

  7. 7.

    Mustard JF, Perry DW, Kinlough-Rathbone RL, Packham MA (1975) Factors responsible for ADP-induced release reaction of human platelets. Am J Physiol 228:1757–1765

  8. 8.

    Packham MA, Mustard JF (2005) Platelet aggregation and adenosine diphosphate/adenosine triphosphate receptors: a historical perspective. Semin Thromb Hemost 31:129–138

  9. 9.

    Cattaneo M, Gachet C, Cazenave J-P, Packham MA (2002) Adenosine diphosphate (ADP) does not induce thromboxane A2 generation in human platelets. Blood 99:3868–3869

  10. 10.

    Cattaneo M, Canciani MT, Lecchi A, Kinlough-Rathbone RL, Packham MA, Mannucci PM, Mustard JF (1990) Released adenosine diphosphate stabilizes thrombin-induced human platelet aggregates. Blood 75:1081–1086

  11. 11.

    Cattaneo M, Akkawat B, Kinlough-Rathbone RL, Packham MA, Cimminiello C, Mannucci PM (1994) Ticlopidine facilitates the deaggregation of human platelets aggregated by thrombin. Thromb Haemost 71:91–94

  12. 12.

    Trumel C, Payrastre B, Plantavid M, Hechler B, Viala C, Presek P, Martinson EA, Cazenave JP, Chap H, Gachet C (1999) A key role of adenosine diphosphate in the irreversible platelet aggregation induced by the PAR1-activating peptide through the late activation of phosphoinositide 3-kinase. Blood 94:4156–4165

  13. 13.

    Cattaneo M, Lecchi A (2007) Inhibition of the platelet P2Y12 receptor for adenosine diphosphate potentiates the antiplatelet effect of prostacyclin. J Thromb Haemost 5:577–582

  14. 14.

    Baurand A, Raboisson P, Freund M, Léon C, Cazenave JP, Bourguignon JJ, Gachet C (2001) Inhibition of platelet function by administration of MRS2179, a P2Y1 receptor antagonist. Eur J Pharmacol 412:213–221

  15. 15.

    Mustard JF, Packham MA (1970) Factors influencing platelet function: adhesion, release and aggregation. Pharmacol Rev 22:97–187

  16. 16.

    MacFarlane DE, Mills DC (1975) The effects of ATP on platelets: evidence against the central role of released ADP in platelet aggregation. Blood 46:309–320

  17. 17.

    Rolf MG, Brearley CA, Mahaut-Smith MP (2001) Platelet shape change evoked by selective activation of P2X1 purinoceptors with alpha, beta-methylene ATP. Thromb Haemost 85:303–308

  18. 18.

    Cattaneo M, Marchese P, Jacobson KA, Ruggeri Z (2002) New insights into the role of P2X1 in platelet function (abstract). Haematologica 87:13–14

  19. 19.

    Cattaneo M, Marchese P, Jacobson KA, Ruggeri ZM (2003) The P2X1 receptor for ATP plays an essential role in platelet aggregation at high shear. Haematol J 4(suppl 2):150, Abstract

  20. 20.

    Oury C, Toth-Zsamboki E, Thys C, Tytgat J, Vermylen J, Hoylaerts MF (2001) The ATP-gated P2X1 ion channel acts as a positive regulator of platelet responses to collagen. Thromb Haemost 86:1264–1271

  21. 21.

    Oury C, Sticker E, Cornelissen H, De Vos R, Vermylen J, Hoylaerts MF (2004) ATP augments von Willebrand factor-dependent shear-induced platelet aggregation through Ca2+-calmodulin and myosin light chain kinase activation. J Biol Chem 279:26266–26273

  22. 22.

    Hechler B, Lenain N, Marchese P, Vial C, Heim V, Freund M, Cazenave JP, Cattaneo M, Ruggeri ZM, Evans R, Gachet C (2003) A role for the fast ATP-gated P2X1 cation channel P2X1 in thrombosis of small arteries in vivo. J Exp Med 198:661–667

  23. 23.

    Oury C, Toth-Zsamboki E, Van Geet C, Thys C, Wei L, Nilius B, Vermylen J, Hoylaerts MF (2000) A natural dominant negative P2X1 receptor due to deletion of a single amino acid residue. J Biol Chem 275:22611–22614

  24. 24.

    Oury C, Lenaerts T, Peerlinck K, Vermylen J, Hoylaerts MF (1999) Congenital deficiency of the phospholipase C coupled platelet P2Y1 receptor leads to a mild bleeding disorder. Thromb Haemost;85(suppl):20 (abs.)

  25. 25.

    Hetherington SL, Singh RK, Lodwick D, Thompson JR, Goodall AH, Samani NJ (2005) Dimorphism in the P2Y1 ADP receptor gene is associated with increased platelet activation response to ADP. Arterioscler Thromb Vasc Biol 25:252–257

  26. 26.

    Cattaneo M, Lecchi A, Randi AM, McGregor JL, Mannucci PM (1992) Identification of a new congenital defect of platelet function characterized by severe impairment of platelet responses to adenosine diphosphate. Blood 80:2787–2796

  27. 27.

    Gachet C, Cattaneo M, Ohlmann P, Hechler B, Lecchi A, Chevalier J, Cassel D, Mannucci PM, Cazenave JP (1995) Purinoceptors on blood platelets: further pharmacological and clinical evidence to suggest the presence of two ADP receptors. Br J Haematol 91:434–444

  28. 28.

    Nurden P, Savi P, Heilmann E, Bihour C, Herbert JM, Maffrand JP, Nurden A (1995) An inherited bleeding disorder linked to a defective interaction between ADP and its receptor on platelets. Its influence on glycoprotein IIb-IIIa complex function. J Clin Invest 95:1612–1622

  29. 29.

    Cattaneo M, Lecchi A, Lombardi R, Gachet C, Zighetti ML (2000) Platelets from a patient heterozygous for the defect of P2CYC receptors for ADP have a secretion defect despite normal thromboxane A2 production and normal granule stores: further evidence that some cases of platelet ‘primary secretion defect’ are heterozygous for a defect of P2CYC receptors. Arterioscler Thromb Vasc Biol 20:E101–E106

  30. 30.

    Shiraga M, Miyata S, Kato H, Kashiwagi H, Honda S, Kurata Y, Tomiyama Y, Kanakura Y (2005) Impaired platelet function in a patients with P2Y12 deficiency caused by a mutation in the translation initiation codon. J Thromb Haemost 3:2315–2323

  31. 31.

    Dawood BB, Daly M, Makris M, Mumford A, Watson S, Wilde J (2009) Identification of a novel homozygous P2Y12 mutation in a patient with a mild platelet-based bleeding disorder [abstract]. J Thromb Haemost; volume 7, supplement 2: Abstract PP-MO-076

  32. 32.

    Cattaneo M, Lombardi R, Zighetti ML, Gachet C, Ohlmann P, Cazenave J-P, Mannucci PM (1997) Deficiency of [33P]2MeS-ADP binding sites on platelets with secretion defect, normal granule stores and normal thromboxane A2 production. Evidence that ADP potentiates platelet secretion independently of the formation of large platelet aggregates and thromboxane A2 production. Thromb Haemost 77:986–990

  33. 33.

    Cattaneo M (2003) Inherited platelet-based bleeding disorders. J Thromb Haemost 1:1628–1636

  34. 34.

    Conley PB, Jurek MM, Vincent D, Lecchi A, Cattaneo M (2001) Unique mutations in the P2Y12 locus of patients with previously described defects in ADP-dependent aggregation [abstract]. Blood 98:43b

  35. 35.

    Fontana G, Ware J, Cattaneo M (2009) Haploinsufficiency of the platelet P2Y12 gene in a family with congenital bleeding diathesis. Haematologica 94:581–584

  36. 36.

    Hollopeter G, Jantzen HM, Vincent D, Li G, England L, Ramakrishnan V, Yang RB, Nurden P, Nurden A, Julius D, Conley PB (2001) Identification of the platelet ADP receptor targeted by antithrombotic drugs. Nature 409:202–207

  37. 37.

    Cattaneo M, Zighetti ML, Lombardi R, Martinez C, Lecchi A, Conley PB, Ware J, Ruggeri ZM (2003) Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding. Proc Natl Acad Sci USA 100:1978–1983

  38. 38.

    Moro S, Hoffmann C, Jacobson KA (1999) Role of the extracellular loops of G protein-coupled receptors in ligand recognition: a molecular modeling study of the human P2Y1 receptor. Biochemistry 38:3498–3507

  39. 39.

    Moro S, Guo D, Camaioni E, Boyer JL, Harden TK, Jacobson KA (1998) Human P2Y1 receptor: molecular modeling and site-directed mutagenesis as tools to identify agonist and antagonist recognition sites. J Med Chem 41:1456–1466

  40. 40.

    Remijn JA, IJsseldijk MJ, Strunk AL, Abbes AP, Engel H, Dikkeschei B, Dompeling EC, de Groot PG, Slingerland RJ (2007) Novel molecular defect in the platelet ADP receptor P2Y12 of a patient with haemorrhagic diathesis. Clin Chem Lab Med 45:187–189

  41. 41.

    Daly ME, Dawood BB, Lester WA, Peake IR, Rodeghiero F, Goodeve AC, Makris M, Wilde JT, Mumford AD, Watson SP, Mundell SJ (2009) Identification and characterization of a novel P2Y 12 variant in a patient diagnosed with type 1 von Willebrand disease in the European MCMDM-1VWD study. Blood 113:4110–4113

  42. 42.

    Savi P, Zachayus JL, Delesque-Touchard N, Labouret C, Hervé C, Uzabiaga MF, Pereillo JM, Culouscou JM, Bono F, Ferrara P, Herbert JM (2006) The active metabolite of Clopidogrel disrupts P2Y12 receptor oligomers and partitions them out of lipid rafts. Proc Natl Acad Sci USA 103:11069–11074

  43. 43.

    Savi P, Pereillo JM, Uzabiaga MF, Combalbert J, Picard C, Maffrand JP, Pascal M, Herbert JM (2000) Identification and biological activity of the active metabolite of clopidogrel. Thromb Haemost 84:891–896

  44. 44.

    Ding Z, Kim S, Dorsam RT, Jin J, Kunapuli SP (2003) Inactivation of the human P2Y12 receptor by thiol reagents requires interaction with both extracellular cysteine residues, Cys17 and Cys270. Blood 101:3908–3914

  45. 45.

    Algaier I, Jakubowski JA, Asai F, von Kügelgen I (2008) Interaction of the active metabolite of prasugrel, R-138727, with cysteine 97 and cysteine 175 of the human P2Y12 receptor. J Thromb Haemost 6:1908–1914

  46. 46.

    Tosetto A, Rodeghiero F, Castaman G, Goodeve A, Federici AB, Batlle J, Meyer D, Fressinaud E, Mazurier C, Goudemand J, Eikenboom J, Schneppenheim R, Budde U, Ingerslev J, Vorlova Z, Habart D, Holmberg L, Lethagen S, Pasi J, Hill F, Peake I (2006) A quantitative analysis of bleeding symptoms in type 1 von Willebrand disease: results from a multicenter European study (MCMDM-1 VWD). J Thromb Haemost 4:766–773

  47. 47.

    Zighetti ML, Carpani G, Sinigaglia E, Cattaneo M (2010) Usefulness of a flow cytometric analysis of intraplatelet VASP phosphorylation for the detection of patients with genetic defects of the platelet P2Y12 receptor for ADP. J Thromb Haemost 8:2332–2334

  48. 48.

    Cattaneo M, Zighetti ML, Lombardi R, Mannucci PM (1994) Role of ADP in platelet aggregation at high shear: studies in a patient with congenital defect of platelet responses to ADP. Br J Haematol 88:826–829

  49. 49.

    Cattaneo M, Mannucci PM (2002) Desmopressin. In: Michelson AD (ed) Platelets. Academic, San Diego, pp 855–866

  50. 50.

    Fontana P, Dupont A, Gandrille S, Bachelot-Loza C, Reny JL, Aiach M, Gaussem P (2003) Adenosine diphosphate-induced platelet aggregation is associated with P2Y12 gene sequence variations in healthy subjects. Circulation 108:989–995

  51. 51.

    Fontana P, Gaussem P, Aiach M, Fiessinger JN, Emmerich J, Reny JL (2003) P2Y12 H2 haplotype is associated with peripheral arterial disease: a case–control study. Circulation 108:2971–2973

  52. 52.

    Cattaneo M (2004) Aspirin and clopidogrel. efficacy, safety, and the issue of drug resistance. Arterioscler Thromb Vasc Biol 24:1980–1987

  53. 53.

    Cattaneo M (2010) New P2Y(12) inhibitors. Circulation 121:171–179

  54. 54.

    von Beckerath N, von Beckerath O, Koch W, Eichinger M, Schomig A, Kastrati A (2005) P2Y12 gene H2 haplotype is not associated with increased adenosine diphosphate-induced platelet aggregation after initiation of clopidogrel therapy with a high loading dose. Blood Coagul Fibrinolysis 16:199–204

  55. 55.

    Angiolillo DJ, Fernandez-Ortiz A, Bernardo E, Ramírez C, Cavallari U, Trabetti E, Sabaté M, Jimenez-Quevedo P, Hernández R, Moreno R, Escaned J, Alfonso F, Bañuelos C, Costa MA, Bass TA, Pignatti PF, Macaya C (2005) Lack of association between the P2Y12 receptor gene polymorphism and platelet response to clopidogrel in patients with coronary artery disease. Thromb Res 116:491–497

  56. 56.

    von Beckerath N, von Beckerath O, Koch W, Eichinger M, Schömig A, Kastrati A (2005) P2Y12 gene H2 haplotype is not associated with increased adenosine diphosphate-induced platelet aggregation after initiation of clopidogrel therapy with a high loading dose. Blood Coagul Fibrinolysis 16:199–204

  57. 57.

    Ziegler S, Schillinger M, Funk M, Felber K, Exner M, Mlekusch W, Sabeti S, Amighi J, Minar E, Brunner M, Müller M, Mannhalter C (2005) Association of a functional polymorphism in the clopidogrel target receptor gene, P2Y12, and the risk for ischemic cerebrovascular events in patients with peripheral artery disease. Stroke 36:1394–1399

  58. 58.

    Smith SM, Judge HM, Peters G, Armstrong M, Fontana P, Gaussem P, Daly ME, Storey RF (2006) Common sequence variations in the P2Y12 and CYP3A5 genes do not explain the variability in the inhibitory effects of clopidogrel therapy. Platelets 17:250–258

  59. 59.

    Cuisset T, Frere C, Quilici J, Morange PE, Saut N, Lambert M, Camoin L, Vague IJ, Bonnet JL, Alessi MC (2007) Role of the T744C polymorphism of the P2Y12 gene on platelet response to a 600-mg loading dose of clopidogrel in 597 patients with non-ST-segment elevation acute coronary syndrome. Thromb Res 120:893–899

  60. 60.

    Rudez G, Bouman HJ, van Werkum JW, Leebeek FW, Kruit A, Ruven HJ, ten Berg JM, de Maat MP, Hackeng CM (2009) Common variation in the platelet receptor P2RY12 gene is associated with residual on-clopidogrel platelet reactivity in patients undergoing elective percutaneous coronary interventions. Circ Cardiovasc Genet 2:515–521

  61. 61.

    Bouman HJ, van Werkum JW, Rudez G, Leebeek FW, Kruit A, Hackeng CM, ten Berg JM, de Maat MP, Ruven HJ (2010) The influence of variation in the P2Y12 receptor gene on in vitro platelet inhibition with the direct P2Y12 antagonist cangrelor. Thromb Haemost 103:379–386

Download references

Author information

Correspondence to Marco Cattaneo.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Cattaneo, M. Molecular defects of the platelet P2 receptors. Purinergic Signalling 7, 333–339 (2011). https://doi.org/10.1007/s11302-011-9217-z

Download citation

Keywords

  • Platelets
  • P2Y12
  • P2 receptors
  • Adenosine diphosphate
  • Platelet function disorders
  • Platelet physiology