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Thromb Haemost 1993; 70(01): 217-223
DOI: 10.1055/s-0038-1646194
State-of-the-Art Lecture
Platelet Signaling
Schattauer GmbH Stuttgart

Signaling through G Proteins and G Protein-coupled Receptors during Platelet Activation

Lawrence F Brass
Departments of Medicine, Pathology and Pharmacology, University of Pennsylvania. Philadelphia, PA, USA
,
James A Hoxie
Departments of Medicine, Pathology and Pharmacology, University of Pennsylvania. Philadelphia, PA, USA
,
David R Manning
Departments of Medicine, Pathology and Pharmacology, University of Pennsylvania. Philadelphia, PA, USA
› Author Affiliations
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Publication History

Publication Date:
03 July 2018 (online)

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Summary

Recent studies have helped to define the early events of signal transduction in platelets. The best-described of these events are those in which heterotrimeric guanine nucleotide binding regulatory proteins (G proteins) mediate the interaction between cell surface receptors for agonists and intracellular second messenger generating enzymes. To date nine different G proteins have been identified in platelets. The:r targets include phospholipases C and A2, and adenylyl cyclase. Efforts to clone the receptors that can couple to these G proteins have been successful for epinephrine, thrombin, TxA2 and platelet activating factor. Each of these is comprised of a single polypeptide with seven transmembrane domains and an extracellular N-terminus. In the case of the thrombin receptor, activation occurs by a novel mechanism in which thrombin cleaves its receptor, creating a new N-terminus that can serve as a tethered ligand. Shortly after activation, thrombin receptors become resistant to re-activation by thrombin. This desensitization, which appears to involve receptor phosphorylation and internalization, provides a potent mechanism for limiting the duration of thrombin-initiated events in platelets and other thrombin-responsive vascular cells.

 

  • References

  • 1 Hirata M, Hayashi Y, Ushikubi F, Nakanishi S, Narumiya S. Cloning and expression of cDNA for a human thromboxane A2 receptor. Nature 1991; 349: 617-620
    CrossrefPubMedGoogle Scholar
  • 2 Honda Z, Nakamura M, Miki I. et al.: Cloning by functional expression of platelet-activating factor receptor from guinea-pig lung. Nature 1991; 349: 342-346
    CrossrefPubMedGoogle Scholar
  • 3 Vu T-KH, Hung DT, Wheaton VI, Coughlin SR. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-1068
    CrossrefPubMedGoogle Scholar
  • 4 Rasmussen UB, Vouret-Craviari V, Jallat S. et al.: cDNA cloning and expression of a hamster α-thrombin receptor coupled to Ca2+ mobilization. FEBS Lett 1991; 288: 123-128
    CrossrefPubMedGoogle Scholar
  • 5 Kobilka BK, Matsui H, Kobilka TS. et al.: Cloning, sequencing, and expression of the gene coding for the human platelet alpha 2-adren-ergic receptor. Science 1987; 238: 650-656
    CrossrefPubMedGoogle Scholar
  • 6 Harmon JT, Jamieson GA. Activation of platelets by alpha-thrombin is a receptor-mediated event. J Biol Chem 1986; 261: 15928-15933
    PubMedGoogle Scholar
  • 7 De Marco L, Mazzucato M, Masotti A, Fenton II JW, Ruggeri ZM. Function of glycoprotein Iba in platelet activation induced by α-thrombin. J Biol Chem 1991; 266: 23776-23783
    PubMedGoogle Scholar
  • 8 Vu T-KH, Wheaton VI, Hung DT, Charo I, Coughlin SR. Domains specifying thrombin-receptor interaction. Nature 1991; 353: 674-677
    CrossrefPubMedGoogle Scholar
  • 9 Scarborough RM, Naughton MA, Teng W. et al.: Tethered ligand agonist peptides. Structural requirements for thrombin receptor activation reveal mechanism of proteolytic unmasking of agonist function. J Biol Chem 1992; 267: 13146-13149
    PubMedGoogle Scholar
  • 10 Vassallo Jr RR, Kieber-Emmons T, Cichowski K, Brass LF. Structure/function relationships in the activation of platelet thrombin receptors by receptor-derived peptides. J Biol Chem 1992; 267: 6081-6085
    PubMedGoogle Scholar
  • 11 Chao BH, Kalkunte S, Maraganore JM, Stone SR. Essential groups in synthetic agonist peptides for activation of the platelet thrombin receptor. Biochemistry 1992; 31: 6175-6178
    CrossrefPubMedGoogle Scholar
  • 12 Sabo T, Gurwitz D, Motola L, Brodt P, Barak R, Elhanaty E. Structure-activity studies of the thrombin receptor activating peptide. Biochem Biophys Res Commun 1992; 188: 604-610
    CrossrefPubMedGoogle Scholar
  • 13 Brass LF, Vassallo Jr RR, Belmonte E, Ahuja M, Cichowski K, Hoxie JA. Structure and function of the human platelet thrombin receptor: studies using monoclonal antibodies against a defined epitope within the receptor N-terminus. J Biol Chem 1992; 267: 13795-13798
    PubMedGoogle Scholar
  • 14 Hung DT, Vu T-KH, Wheaton VI, Ishii K, Coughlin SR. Cloned platelet thrombin receptor is necessary for thrombin-induced platelet activation. J Clin Invest 1992; 89: 1350-1353
    CrossrefPubMedGoogle Scholar
  • 15 Brass LF, Manning DR, Williams A, Woolkalis MJ, Poncz M. Receptor and G protein-mediated responses to thrombin in HEL cells. J Biol Chem 1991; 266: 958-965
    PubMedGoogle Scholar
  • 16 Brass LF. Homologous desensitization of HEL cell thrombin receptors: distinguishable roles for proteolysis and phosphorylation. J Biol Chem 1992; 267: 6044-6050
    PubMedGoogle Scholar
  • 17 Hoxie JA, Ahuja M, Belmonte E, Pizarro S, Parton RG, Brass LF. Internalization and recycling of activated thrombin receptors. J Biol Chem, in press.
    PubMedGoogle Scholar
  • 18 Rittenhouse SE, Sasson JP. Mass changes in myoinositol trisphos-phate in human platelets stimulated by thrombin. Inhibitory effects of phorbol ester. J Biol Chem 1985; 260: 8657-8660
    PubMedGoogle Scholar
  • 19 Watson SP, Lapetina EG. 1,2-Diacylglycerol and phorbol ester inhibit agonist-induced formation of inositol phosphates in human platelets: possible implications for negative feedback regulation of inositol phospholipid hydrolysis. Proc Natl Acad Sci USA 1985; 82: 2623-2626
    CrossrefPubMedGoogle Scholar
  • 20 Onorato JJ, Palczewski K, Regan JW, Caron MG, Lefkowitz RJ, Benovic JL. Role of acidic amino acids in peptide substrates of the beta-adrenergic receptor kinase and rhodopsin kinase. Biochemistry 1991; 30: 5118-5125
    CrossrefPubMedGoogle Scholar
  • 21 Golden A, Brugge JS, Shattil SJ. Role of platelet membrane glycoprotein Ilb-IIIa in agonist-induced tyrosine phosphorylation of platelet proteins. J Cell Biol 1990; 111: 3117-3127
    CrossrefPubMedGoogle Scholar
  • 22 Jelsema CL, Axelrod J. Stimulation of phospholipase A2 activity in bovine rod outer segments by the beta gamma subunits of transducin and its inhibition by the alpha subunit. Proc Natl Acad Sci USA 1987; 84: 3623-3627
    CrossrefPubMedGoogle Scholar
  • 23 Burch RM, Luini A, Axelrod J. Phospholipase A2 and phospholipase C are activated by distinct GTP-binding proteins in response to alpha 1-adrenergic stimulation in FRTL5 thyroid cells. Proc Natl Acad Sci USA 1986; 83: 7201-7205
    CrossrefPubMedGoogle Scholar
  • 24 Birnbaumer L. Receptor-to-effector signaling through G proteins: Roles for βgamma dimers as well as α subunits. Cell 1992; 71: 1069-1072
    CrossrefPubMedGoogle Scholar
  • 25 Taussig R, Quarmby LM, Gilman AG. Regulation of purified type I and type II adenylylcyclases by G protein βγ subunits. J Biol Chem 1993; 268: 9-12
    PubMedGoogle Scholar
  • 26 Blank JL, Brattain KA, Exton JH. Activation of cytosolic phospho-inositide phospholipase C by G-protein βγ subunits. J Biol Chem 1992; 267: 23069-23075
    PubMedGoogle Scholar
  • 27 Katz A, Wu D, Simon MI. Subunits bgamma of heterotrimeric G protein activate (32 isoform of phospholipase C. Nature 1992; 360: 686-689
    CrossrefPubMedGoogle Scholar
  • 28 Camps M, Carozzi A, Schnabel P, Scheer A, Parker PJ, Gierschik P. Isozyme-selective stimulation of phospholipase C-β2 by G protein (Bgamma-subunits. Nature 1992; 360: 684-686
    CrossrefPubMedGoogle Scholar
  • 29 Ercolani L, Stow JL, Boyle JF, et al. Membrane localization of the pertussis toxin-sensitive G protein subunits αi2 and αi3 and expression of a metallothionein-αi2 fusion gene in LLC-PK1 cells. Proc Natl Acad Sci USA 1990; 87: 4635-4639
    CrossrefPubMedGoogle Scholar
  • 30 Lewis JM, Woolkalis MJ, Gerton GL, Smith RM, Jarett L, Manning DR. Subcellular distribution of the α subunit(s) of Gi: visualization by immunofluorescent and immunogold labeling. Cell Reg 1991; 2: 1097-1113
    PubMedGoogle Scholar
  • 31 Strathmann MP, Simon MI. Gαl2 and Gαl3 subunits define a fourth class of G protein a subunits. Proc Natl Acad Sci USA 1991; 88: 5582-5586
    CrossrefPubMedGoogle Scholar
  • 32 Williams A, Woolkalis MJ, Poncz M, Manning DR, Gewirtz A, Brass LF. Identification of the pertussis toxin-sensitive G proteins in platelets, megakaryocytes and HEL cells. Blood 1990; 76: 721-730
    PubMedGoogle Scholar
  • 33 Brass LF, Laposata M, Banga HS, Rittenhouse SE. Regulation of the phosphoinositide hydrolysis pathway in thrombin-stimulated platelets by a pertussis toxin-sensitive guanine nucleotide-binding protein. Evaluation of its contribution to platelet activation and comparisons with the adenylate cyclase inhibitory protein, Gi. J Biol Chem 1986; 261: 16838-16847
    PubMedGoogle Scholar
  • 34 Brass LF, Shaller CC, Belmonte EJ. Inositol 1,4,5-triphosphate-induced granule secretion in platelets. Evidence that the activation of phospholipase C mediated by platelet thromboxane receptors involves a guanine nucleotide binding protein-dependent mechanism distinct from that of thrombin. J Clin Invest 1987; 79: 1269-1275
    CrossrefPubMedGoogle Scholar
  • 35 Brass LF, Woolkalis MJ, Manning DR. Interactions in platelets between G proteins and the agonists that stimulate phospholipase C and inhibit adenylyl cyclase. J Biol Chem 1988; 263: 5348-5355
    PubMedGoogle Scholar
  • 36 Strathmann M, Simon MI. G protein diversity: a distinct class of a subunits is present in vertebrates and invertebrates. Proc Natl Acad Sci USA 1990; 87: 9113-9117
    CrossrefPubMedGoogle Scholar
  • 37 Wu D, Ho Lee C, Goo Rhee S, Simon MI. Activation of phospholipase C by the α subunits of the Gq and G11 proteins in transfected Cos-7 cells. J Biol Chem 1992; 267: 1811-1817
    PubMedGoogle Scholar
  • 38 Berstein G, Blank JL, Smrcka AV, et al. Reconstitution of agonist-stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis using purified ml muscarinic receptor, Gq/ll, and phospholipase C-β1. J Biol Chem 1992; 267: 8081-8088
    PubMedGoogle Scholar
  • 39 Martin TFJ, Lewis JE, Kowalchyk JA. Phospholipase C-β1 is regulated by a pertussis toxin-insensitive G-protein. Biochem J 1991; 280: 753-760
    CrossrefPubMedGoogle Scholar
  • 40 Shenker A, Goldsmith P, Unson CG, Spiegel AM. The G protein coupled to the thomboxane A2 receptor in human platelets is a member of the novel Gq family. J Biol Chem 1991; 266: 9309-9313
    PubMedGoogle Scholar
  • 41 Smrcka AV, Hepler JR, Brown KO, Sternweis PC. Regulation of polyphosphoinositide-specific phospholipase C activity by purified Gq . Science 1991; 251: 804-807
    CrossrefPubMedGoogle Scholar
  • 42 Taylor SJ, Exton JH. Two α subunits of the Gq class of G proteins stimulate phosphoinositide phospholipase C-β1 activity. FEBS Lett 1991; 286: 214-216
    CrossrefPubMedGoogle Scholar
  • 43 Taylor SJ, Chae HZ, Rhee SG, Exton JH. Activation of the β1 isozyme of phospholipase C by α subunits of the Gq class of G proteins. Nature 1991; 350: 516-518
    CrossrefPubMedGoogle Scholar
  • 44 Blank JL, Ross AH, Exton JH. Purification and characterization of two G-proteins that activate the β1 isozyme of phosphoinositide-spe-cific phospholipase C. Identification as members of the Gq class. J Biol Chem 1991; 266: 18206-18216
    PubMedGoogle Scholar
  • 45 Berridge MJ. Inositol trisphosphate and calcium signalling. Nature 1993; 361: 315-325
    CrossrefPubMedGoogle Scholar
  • 46 Kim D, Lewis DL, Graziadei L, Neer EJ, Bar-Sagi D, Clapham DE. G-protein βgamma-subunits activate the cardiac muscarinic K+-channel via phospholipase A2 . Nature 1989; 337: 557-560
    CrossrefPubMedGoogle Scholar
  • 47 Carlson K, Brass LF, Manning DR. Thrombin and phorbol esters cause the selective phosphorylation of a G protein other than Gi in human platelets. J Biol Chem 1989; 264: 13298-13305
    PubMedGoogle Scholar
  • 48 Gagnon AW, Manning DR, Catani L, Gewirtz A, Poncz M, Brass LF. Identification of Gzα as a pertussis toxin-insensitive G protein in human platelets and megakaryocytes. Blood 1991; 78: 1247-1253
    PubMedGoogle Scholar
  • 49 Lounsbury KM, Casey PJ, Brass LF, Manning DR. Phosphorylation of Gz in human platelets: selectivity and site of modification. J Biol Chem 1991; 266: 22051-22056
    PubMedGoogle Scholar
  • 50 Lounsbury KM, Schlegel B, Poncz M, Brass LF, Manning DR. Analysis of Gzα by site-directed mutagenesis: sites and specificity of protein kinase C-dependent phosphorylation. J Biol Chem 1993; 268: 3494-3498
    PubMedGoogle Scholar
  • 51 Amatruda III TT, Steele DA, Slepak VZ, Simon MI. Gα16, a G protein α subunit specifically expressed in hematopoietic cells. Proc Natl Acad Sci USA 1991; 88: 5587-5591
    CrossrefPubMedGoogle Scholar
  • 52 Lounsbury KM, Brass LF, Manning DR. Phosphorylation of Gzα in human platelets: proximity to the amino-terminus of the subunit. J Cell Biol 1990; 111: 334a
    PubMedGoogle Scholar
  • 53 Smith SK, Limbird LE. Evidence that human platelet alpha-adrener-gic receptors coupled to inhibition of adenylate cyclase are not associated with the subunit of adenylate cyclase ADP-ribosylated by cholera toxin. J Biol Chem 1982; 257: 10471-10478
    PubMedGoogle Scholar