Home LiteratureArticle Details
PMID: 9020890 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Ras activation in platelets after stimulation of the thrombin receptor, thromboxane A2 receptor or protein kinase C.

The Biochemical journal ·Vol. 321 ( Pt 2) ·1997-01-15 ·Pages 525-30

Shock DD, He K, Wencel-Drake JD, Parise LV

Abstract

Several reports have indicated that the small G-protein Ras is not present immunologically in platelets. However, here we report the identification of Ras in platelets by immunoprecipitation with the Ras-specific monoclonal antibodies Y13-259 or Y13-238, followed by Western blotting. The presence of Ras was not due to contamination of samples with erythrocytes or leucocytes. Immunofluorescence studies indicated that Ras was present in a peripheral rim pattern in fixed, permeabilized platelets, suggesting an intracellular, plasma membrane location. Activation of platelets with the thrombin receptor peptide42-50, the prostaglandin H2/thromboxane A2 mimetic U46619 or phorbol 12-myristate 13-acetate induced a rapid increase in GTP-bound, activated Ras. In each case, this increase was inhibited by the protein kinase C (PKC) inhibitor bisindolylmaleimide GF 109203X, suggesting that Ras is activated downstream of PKC in platelets. Thus the activation of Ras in platelets by agonists will now allow consideration of multiple potential Ras-dependent signal transduction pathways in platelet activation processes.

MeSH Terms
Blood Platelets/enzymology,metabolism Densitometry GTP-Binding Proteins/blood Humans Precipitin Tests Protein Kinase C/blood,metabolism Proto-Oncogene Proteins p21(ras)/blood,metabolism Receptors, Thrombin/blood,metabolism Receptors, Thromboxane/blood,metabolism
Chemicals
Receptors, Thrombin Receptors, Thromboxane Protein Kinase C GTP-Binding Proteins HRAS protein, human Proto-Oncogene Proteins p21(ras)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Shock D D
Department of Pharmacology, University of North Carolina at Chapel Hill 27599-7365, USA.
He K
Wencel-Drake J D
Parise L V
References (45)
45 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Integrin activation by R-ras.
    Cell. 1996 Apr 5;85(1):61-9 PMID: 8620538
  3. Localization of the src gene product of the Harvey strain of MSV to plasma membrane of transformed cells by electron microscopic immunocytochemistry.
    Cell. 1980 Apr;19(4):1005-14 PMID: 6247068
  4. Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family.
    J Virol. 1982 Jul;43(1):294-304 PMID: 6287003
  5. ras gene alterations and enhanced levels of ras p21 expression in a spectrum of benign and malignant human mammary tissues.
    Lab Invest. 1986 Dec;55(6):603-15 PMID: 2431221
  6. Expression of p21ras in normal and malignant human tissues: lack of association with proliferation and malignancy.
    Proc Natl Acad Sci U S A. 1987 May;84(10):3234-8 PMID: 3554234
  7. Gn-proteins are distinct from ras p21 and other known low molecular mass GTP-binding proteins in the platelet.
    FEBS Lett. 1988 Sep 12;237(1-2):168-72 PMID: 3139450
  8. Small molecular weight GTP-binding proteins in human platelet membranes. Purification and characterization of a novel GTP-binding protein with a molecular weight of 22,000.
    J Biol Chem. 1989 Jan 25;264(3):1877-81 PMID: 2492285
  9. A ras-related protein is phosphorylated and translocated by agonists that increase cAMP levels in human platelets.
    Proc Natl Acad Sci U S A. 1989 May;86(9):3131-4 PMID: 2470091
  10. Thrombolamban, the 22-kDa platelet substrate of cyclic AMP-dependent protein kinase, is immunologically homologous with the Ras family of GTP-binding proteins.
    Proc Natl Acad Sci U S A. 1990 Jan;87(2):758-62 PMID: 1689052
  11. Multiple small molecular weight guanine nucleotide-binding proteins in human erythrocyte membranes.
    Biochem Biophys Res Commun. 1990 Feb 14;166(3):1398-405 PMID: 2106317
  12. Human platelets form 3-phosphorylated phosphoinositides in response to alpha-thrombin, U46619, or GTP gamma S.
    J Biol Chem. 1990 Apr 5;265(10):5345-8 PMID: 2156815
  13. ras p21 and GAP inhibit coupling of muscarinic receptors to atrial K+ channels.
    Cell. 1990 Jun 1;61(5):769-76 PMID: 2188736
  14. Stimulation of p21ras upon T-cell activation.
    Nature. 1990 Aug 23;346(6286):719-23 PMID: 2201921
  15. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation.
    Cell. 1991 Mar 22;64(6):1057-68 PMID: 1672265
  16. Phosphorylation of human platelet glycoprotein IIIa (GPIIIa). Dissociation from fibrinogen receptor activation and phosphorylation of GPIIIa in vitro.
    J Biol Chem. 1991 Aug 5;266(22):14663-9 PMID: 1650365
  17. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C.
    J Biol Chem. 1991 Aug 25;266(24):15771-81 PMID: 1874734
  18. Ras is essential for nerve growth factor- and phorbol ester-induced tyrosine phosphorylation of MAP kinases.
    Cell. 1992 Mar 20;68(6):1031-40 PMID: 1312392
  19. ras mediates nerve growth factor receptor modulation of three signal-transducing protein kinases: MAP kinase, Raf-1, and RSK.
    Cell. 1992 Mar 20;68(6):1041-50 PMID: 1312393
  20. Involvement of p21ras in activation of extracellular signal-regulated kinase 2.
    Nature. 1992 Jun 18;357(6379):602-4 PMID: 1608472
  21. Role of rap1B and p21ras GTPase-activating protein in the regulation of phospholipase C-gamma 1 in human platelets.
    Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7796-800 PMID: 1323853
  22. The cloned platelet thrombin receptor couples to at least two distinct effectors to stimulate phosphoinositide hydrolysis and inhibit adenylyl cyclase.
    J Biol Chem. 1992 Oct 15;267(29):20831-4 PMID: 1328213
  23. A rho gene product in human blood platelets. II. Effects of the ADP-ribosylation by botulinum C3 ADP-ribosyltransferase on platelet aggregation.
    J Biol Chem. 1992 Oct 15;267(29):20921-6 PMID: 1400407
  24. Pertussis toxin-sensitive activation of p21ras by G protein-coupled receptor agonists in fibroblasts.
    Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1257-61 PMID: 7679495
  25. Activation of myelin basic protein and S6 peptide kinases in phorbol ester- and PAF-treated sheep platelets.
    Biochim Biophys Acta. 1993 Apr 16;1176(3):287-98 PMID: 8385998
  26. Regulation of vascular integrins.
    Blood. 1993 Jun 1;81(11):2827-43 PMID: 8499621
  27. The effects of genistein on platelet function are due to thromboxane receptor antagonism rather than inhibition of tyrosine kinase.
    Prostaglandins Leukot Essent Fatty Acids. 1993 May;48(5):379-84 PMID: 8321874
  28. A requirement for Ras protein function in thrombin-stimulated mitogenesis in astrocytoma cells.
    J Biol Chem. 1993 Sep 15;268(26):19411-5 PMID: 8396137
  29. p42 mitogen-activated protein kinase and p90 ribosomal S6 kinase are selectively phosphorylated and activated during thrombin-induced platelet activation and aggregation.
    Mol Cell Biol. 1994 Jan;14(1):463-72 PMID: 8264614
  30. Different pathways for control of Na+/H+ exchange via activation of the thrombin receptor.
    Biochem J. 1994 Jan 1;297 ( Pt 1):47-52 PMID: 8280110
  31. Tyrosine phosphorylation and activation of mitogen-activated protein kinases by thrombin in human platelets: possible involvement in late arachidonic acid release.
    Biochem Biophys Res Commun. 1994 Jan 28;198(2):497-503 PMID: 8297360
  32. Interaction of Ras and Raf in intact mammalian cells upon extracellular stimulation.
    J Biol Chem. 1994 Feb 11;269(6):3913-6 PMID: 8307946
  33. Phosphatidylinositol-3-OH kinase as a direct target of Ras.
    Nature. 1994 Aug 18;370(6490):527-32 PMID: 8052307
  34. Direct evidence that Gi-coupled receptor stimulation of mitogen-activated protein kinase is mediated by G beta gamma activation of p21ras.
    Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12706-10 PMID: 7809106
  35. The role of p21ras in receptor tyrosine kinase signalling.
    Biochim Biophys Acta. 1994 Dec 30;1198(2-3):131-47 PMID: 7819271
  36. Sequestration of a G-protein beta gamma subunit or ADP-ribosylation of Rho can inhibit thrombin-induced activation of platelet phosphoinositide 3-kinases.
    J Biol Chem. 1995 Mar 24;270(12):6589-94 PMID: 7896797
  37. An essential role for Rac in Ras transformation.
    Nature. 1995 Mar 30;374(6521):457-9 PMID: 7700355
  38. Ras-dependent induction of cellular responses by constitutively active phosphatidylinositol-3 kinase.
    Science. 1995 Apr 7;268(5207):100-2 PMID: 7701328
  39. Phosphoinositide 3-kinase inhibition spares actin assembly in activating platelets but reverses platelet aggregation.
    J Biol Chem. 1995 May 12;270(19):11358-66 PMID: 7744773
  40. Gq/11 communicates with thromboxane A2 receptors in human astrocytoma cells, rabbit astrocytes and human platelets.
    Res Commun Mol Pathol Pharmacol. 1995 Mar;87(3):243-51 PMID: 7620818
  41. Effects of thrombin and the thrombin receptor activating peptide, SFLLRN, on redistribution of platelet alpha-granule contents are similar and independent of the extent of thromboxane formation.
    Exp Mol Pathol. 1995 Feb;62(1):63-72 PMID: 7556592
  42. Analysis of Ras protein expression in mammalian cells.
    Methods Enzymol. 1995;255:195-220 PMID: 8524103
  43. Activation of Rac1, RhoA, and mitogen-activated protein kinases is required for Ras transformation.
    Mol Cell Biol. 1995 Nov;15(11):6443-53 PMID: 7565796
  44. Overexpression of the Ras-related TC21/R-Ras2 protein may contribute to the development of human breast cancers.
    Oncogene. 1996 Jan 4;12(1):169-76 PMID: 8552388
  45. Human platelets possess an inducible and saturable receptor specific for fibrinogen.
    J Biol Chem. 1979 Jun 25;254(12):5357-63 PMID: 447655
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3210525
Published
1997-01-15
Pages
525-30
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1218100
Subset
IM
Grants
NHLBI NIH HHS · 1-PO1-HL45100 · United States
External Links
PubMed source DOI: 10.1042/bj3210525 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2