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Thromb Haemost 2012; 108(01): 119-132
DOI: 10.1160/TH11-10-0749
Platelets and Blood Cells
Schattauer GmbH

Farnesyl pyrophosphate is an endogenous antagonist to ADP-stimulated P2Y12 receptor-mediated platelet aggregation

Carl Högberg
1   Department of Cardiology, Lund University, Lund, Sweden
,
Olof Gidlöf
1   Department of Cardiology, Lund University, Lund, Sweden
,
Francesca Deflorian
2   National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA
,
Kenneth A. Jacobson
2   National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA
,
Aliaa Abdelrahman
3   PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany
,
Christa E. Müller
3   PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, Bonn, Germany
,
Björn Olde
1   Department of Cardiology, Lund University, Lund, Sweden
,
David Erlinge
1   Department of Cardiology, Lund University, Lund, Sweden
› Author Affiliations
Further Information

Publication History

Received: 17 November 2011

Accepted after major revision: 20 April 2012

Publication Date:
22 November 2017 (online)

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Summary

Farnesyl pyrophosphate (FPP) is an intermediate in cholesterol biosynthesis, and it has also been reported to activate platelet LPA (lysophos-phatidic acid) receptors. The aim of this study was to investigate the role of extracellular FPP in platelet aggregation. Human platelets were studied with light transmission aggregometry, flow cytometry and [35S]GTPγS binding assays. As shown previously, FPP could potentiate LPA-stimulated shape change. Surprisingly, FPP also acted as a selective insurmountable antagonist to ADP-induced platelet aggregation. FPP inhibited ADP-induced expression of P-selectin and the activated glycoprotein (Gp)IIb/IIIa receptor. FPP blocked ADP-induced inhibition of cAMP accumulation and [35S]GTPγS binding in platelets. In Chinese hamster ovary cells expressing the P2Y12 receptor, FPP caused a right-ward shift of the [35S]GTPγS binding curve. In Sf9 insect cells expressing the human P2Y12 receptor, FPP showed a concentration-dependent, although incomplete inhibition of [3H]PSB-0413 binding. Docking of FPP in a P2Y12 receptor model revealed molecular similarities with ADP and a good fit into the binding pocket for ADP. In conclusion, FPP is an insurmountable antagonist of ADP-induced platelet aggregation mediated by the P2Y12 receptor. It could be an endogenous antithrombotic factor modulating the strong platelet aggregatory effects of ADP in a manner similar to the use of clopidogrel, prasugrel or ticagrelor in the treatment of ischaemic heart disease.

Keywords

ADP receptors - platelet pharmacology - platelet physiology
 

  • References

  • 1 Goldstein JL, Brown MS. Regulation of the mevalonate pathway. Nature 1990; 343: 425-430.
    CrossrefPubMedGoogle Scholar
  • 2 McTaggart SJ. Isoprenylated proteins. Cell Mol Life Sci 2006; 63: 255-267.
    CrossrefPubMedGoogle Scholar
  • 3 Saisho Y, Morimoto A, Umeda T. Determination of farnesyl pyrophosphate in dog and human plasma by high-performance liquid chromatography with fluorescence detection. Analyt Biochem 1997; 252: 89-95.
    CrossrefPubMedGoogle Scholar
  • 4 Williams JR, Khandoga AL, Goyal P. et al. Unique ligand selectivity of the GPR92/LPA5 lysophosphatidate receptor indicates role in human platelet activation. J Biol Chem 2009; 284: 17304-17319.
    CrossrefPubMedGoogle Scholar
  • 5 Amisten S, Braun OO, Bengtsson A. et al. Gene expression profiling for the identification of G-protein coupled receptors in human platelets. Thromb Res 2008; 122: 47-57.
    CrossrefPubMedGoogle Scholar
  • 6 Khandoga AL, Fujiwara Y, Goyal P. et al. Lysophosphatidic acid-induced platelet shape change revealed through LPA(1–5) receptor-selective probes and albumin. Platelets 2008; 19: 415-427.
    CrossrefPubMedGoogle Scholar
  • 7 Hogberg C, Erlinge D, Braun OO. Mild hypothermia does not attenuate platelet aggregation and may even increase ADP-stimulated platelet aggregation after clopidogrel treatment. Thromb J 2009; 07: 2.
    PubMedGoogle Scholar
  • 8 Cazenave JP, Ohlmann P, Cassel D. et al. Preparation of washed platelet suspensions from human and rodent blood. Methods Mol Biol 2004; 272: 13-28.
    PubMedGoogle Scholar
  • 9 Liu EC, Abell LM. Development and validation of a platelet calcium flux assay using a fluorescent imaging plate reader. Analyt Biochem 2006; 357: 216-224.
    CrossrefPubMedGoogle Scholar
  • 10 Vasiljev KS, Uri A, Laitinen JT. 2-Alkylthio-substituted platelet P2Y12 receptor antagonists reveal pharmacological identity between the rat brain Gi-linked ADP receptors and P2Y12. Neuropharmacol 2003; 45: 145-154.
    CrossrefPubMedGoogle Scholar
  • 11 vanGiezen JJ, Nilsson L, Berntsson P. et al. Ticagrelor binds to human P2Y(12) independently from ADP but antagonizes ADP-induced receptor signaling and platelet aggregation. J Thromb Haemost 2009; 07: 1556-1565.
    CrossrefPubMedGoogle Scholar
  • 12 von Kugelgen I, Schiedel AC, Hoffmann K. et al. Cloning and functional expression of a novel Gi protein-coupled receptor for adenine from mouse brain. Mol Pharmacol 2008; 73: 469-477.
    PubMedGoogle Scholar
  • 13 El-Tayeb A, Griessmeier KJ, Muller CE. Synthesis and preliminary evaluation of [3H]PSB-0413, a selective antagonist radioligand for platelet P2Y12 receptors. Bioorg Med Chem Lett 2005; 15: 5450-5452.
    CrossrefPubMedGoogle Scholar
  • 14 Jaakola VP, Griffith MT, Hanson MA. et al. The 2.6 angstrom crystal structure of a human A2A adenosine receptor bound to an antagonist. Science 2008; 322: 1211-1217.
    CrossrefPubMedGoogle Scholar
  • 15 Berman HM, Westbrook J, Feng Z. et al. The Protein Data Bank. Nucleic Acids Res 2000; 28: 235-242.
    CrossrefPubMedGoogle Scholar
  • 16 Sherman W, Day T, Jacobson M P. et al. Novel procedure for modeling ligand/receptor induced fit effects. J Med Chem 2006; 49: 534-553.
    CrossrefPubMedGoogle Scholar
  • 17 Cattaneo M, Zighetti ML, Lombardi R. et al. Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding. Proc Natl Acad Sci USA 2003; 100: 1978-1983.
    CrossrefPubMedGoogle Scholar
  • 18 Hoffmann K, Sixel U, Di Pasquale F. et al. Involvement of basic amino acid residues in transmembrane regions 6 and 7 in agonist and antagonist recognition of the human platelet P2Y(12)-receptor. Biochem Pharmacol 2008; 76: 1201-1213.
    CrossrefPubMedGoogle Scholar
  • 19 Costanzi S, Mamedova L, Gao ZG. et al. Architecture of P2Y nucleotide receptors: structural comparison based on sequence analysis, mutagenesis, and homology modeling. J Med Chem 2004; 47: 5393-5404.
    CrossrefPubMedGoogle Scholar
  • 20 Ivanov AA, Costanzi S, Jacobson KA. Defining the nucleotide binding sites of P2Y receptors using rhodopsin-based homology modeling. J Comp Aid Mol Design 2006; 20: 417-426.
    CrossrefPubMedGoogle Scholar
  • 21 Deflorian F, Jacobson KA. Comparison of three GPCR structural templates for modeling of the P2Y12 nucleotide receptor. J Comput Aided Mol Des 2011; 25: 329-338.
    CrossrefPubMedGoogle Scholar
  • 22 Oh DY, Yoon JM, Moon MJ. et al. Identification of farnesyl pyrophosphate and N-arachidonylglycine as endogenous ligands for GPR92. J Biol Chem 2008; 283: 21054-21064.
    CrossrefPubMedGoogle Scholar
  • 23 Cui G, Merz Jr KM. Computational studies of the farnesyltransferase ternary complex part II: the conformational activation of farnesyldiphosphate. Biochemistry 2007; 46: 12375-12381.
    CrossrefPubMedGoogle Scholar
  • 24 Zahn TJ, Ksebati MB, Gibbs RA. Synthesis and conformational analysis of di13C-labeled farnesyl diphosphate analogs. Tetrahedron Lett 1998; 39: 3991-3994.
    CrossrefPubMedGoogle Scholar
  • 25 Zahn TJ, Eilers M. Evaluation of Isoprenoid Conformation in Solution and in the Active Site of Protein-Farnesyl. J Am Chem Soc 2000; 122: 7153-7165.
    CrossrefPubMedGoogle Scholar
  • 26 Lee M, Choi S, Hallden G. et al. P2Y5 is a G{alpha}i, G{alpha}12/13 G Protein Coupled Receptor Activated by Lysophosphatidic Acid that Reduces Intestinal Cell Adhesion. Am J Physiol 2009; 297: G641-654.
    PubMedGoogle Scholar
  • 27 Liliom K, Tsukahara T, Tsukahara R. et al. Farnesyl phosphates are endogenous ligands of lysophosphatidic acid receptors: inhibition of LPA GPCR and activation of PPARs. Biochim Biophys Acta 2006; 1761: 1506-1514.
    CrossrefPubMedGoogle Scholar
  • 28 Bang S, Yoo S, Yang TJ. et al. Farnesyl pyrophosphate is a novel pain-producing molecule via specific activation of TRPV3. J Biol Chem 2010; 285: 19362-19371.
    CrossrefPubMedGoogle Scholar
  • 29 Gan X, Kaplan R, Menke JG. et al. Dual mechanisms of ABCA1 regulation by geranylgeranyl pyrophosphate. J Biol Chem 2001; 276: 48702-48708.
    CrossrefPubMedGoogle Scholar
  • 30 Fredriksson R, Lagerstrom MC, Lundin LG. et al. The G-protein-coupled receptors in the human genome form five main families. Phylogenetic analysis, paralogon groups, and fingerprints. Mol Pharmacol 2003; 63: 1256-1272.
    CrossrefPubMedGoogle Scholar
  • 31 Surgand JS, Rodrigo J, Kellenberger E. et al. A chemogenomic analysis of the transmembrane binding cavity of human G-protein-coupled receptors. Proteins 2006; 62: 509-538.
    PubMedGoogle Scholar
  • 32 Schwartz GG, Olsson AG, Ezekowitz MD. et al. Effects of atorvastatin on early recurrent ischemic events in acute coronary syndromes: the MIRACL study: a randomized controlled trial. J Am Med Assoc 2001; 285: 1711-1718.
    CrossrefPubMedGoogle Scholar
  • 33 Patti G, Pasceri V, Colonna G. et al. Atorvastatin pretreatment improves outcomes in patients with acute coronary syndromes undergoing early percutaneous coronary intervention: results of the ARMYDA-ACS randomized trial. J Am Coll Cardiol 2007; 49: 1272-1278.
    CrossrefPubMedGoogle Scholar
  • 34 Labios M, Martinez M, Gabriel F. et al. Effect of atorvastatin upon platelet activation in hypercholesterolemia, evaluated by flow cytometry. Thromb Res 2005; 115: 263-270.
    CrossrefPubMedGoogle Scholar