Thrombin-induced translocation of GLUT3 glucose transporters in human platelets

Lynn R. Sorbara, Theresa M. Davies-Hill, Ellen M. Koehler-Stec, Susan J. Vannucci, McDonald K. Horne, Ian A. Simpson

Research output: Contribution to journalArticle

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Abstract

Platelets derive most of their energy from anaerobic glycolysis; during activation this requirement rises approx. 3-fold. To accommodate the high glucose flux, platelets express extremely high concentrations (155 ± 18 pmol/mg of membrane protein) of the most active glucose transporter isoform, GLUT3. Thrombin, a potent platelet activator, was found to stimulate 2-deoxyglucose transport activity 3-5-fold within 10 min at 25°C, with a half-time of 1-2 min. To determine the mechanism underlying the increase in glucose transport activity, an impermeant photolabel, [2-3H]2N-4-(1-azi-2,2,2-trifluoethyl)benzoyl-1,3,-bis-(D-mannose-4-ylozy)-2 -propylamine, was used to covalently bind glucose transporters accessible to the extracellular milieu. In response to thrombin, the level of transporter labelling increased 2.7-fold with a half-time of 1-2 min. This suggests a translocation of GLUT3 transporters from an intracellular site to the plasma membrane in a manner analogous to that seen for the translocation of GLUT4 in insulin-stimulated rat adipose cells. To investigate whether a similar signalling pathway was involved in both systems, platelets and adipose cells were exposed to staurosporin and wortmannin, two inhibitors of GLUT4 translocation in adipose cells. Thrombin stimulation of glucose transport activity in platelets was more sensitive to staurosporin inhibition than was insulin-stimulated transport activity in adipose cells, but it was totally insensitive to wortmannin. This indicates that the GLUT3 translocation in platelets is mediated by a protein kinase C not by a phosphatidylinositol 3-kinase mechanism. In support of this contention, the phorbol ester PMA, which specifically activates protein kinase C, fully stimulated glucose transport activity in platelets and was equally sensitive to inhibition by staurosporin. This study provides a cellular mechanism by which platelets enhance their capacity to import glucose to fulfil the increased energy demands associated with activation.

Original languageEnglish (US)
Pages (from-to)511-516
Number of pages6
JournalBiochemical Journal
Volume328
Issue number2
DOIs
StatePublished - Dec 1 1997

Fingerprint

Facilitative Glucose Transport Proteins
Platelets
Thrombin
Blood Platelets
Glucose
Protein Kinase C
Chemical activation
Phosphatidylinositol 3-Kinase
Insulin
Deoxyglucose
Phorbol Esters
Glycolysis
Cell membranes
Mannose
Labeling
Rats
Protein Isoforms
Membrane Proteins
Cell Membrane
Fluxes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sorbara, L. R., Davies-Hill, T. M., Koehler-Stec, E. M., Vannucci, S. J., Horne, M. K., & Simpson, I. A. (1997). Thrombin-induced translocation of GLUT3 glucose transporters in human platelets. Biochemical Journal, 328(2), 511-516. https://doi.org/10.1042/bj3280511
Sorbara, Lynn R. ; Davies-Hill, Theresa M. ; Koehler-Stec, Ellen M. ; Vannucci, Susan J. ; Horne, McDonald K. ; Simpson, Ian A. / Thrombin-induced translocation of GLUT3 glucose transporters in human platelets. In: Biochemical Journal. 1997 ; Vol. 328, No. 2. pp. 511-516.
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Sorbara, LR, Davies-Hill, TM, Koehler-Stec, EM, Vannucci, SJ, Horne, MK & Simpson, IA 1997, 'Thrombin-induced translocation of GLUT3 glucose transporters in human platelets', Biochemical Journal, vol. 328, no. 2, pp. 511-516. https://doi.org/10.1042/bj3280511

Thrombin-induced translocation of GLUT3 glucose transporters in human platelets. / Sorbara, Lynn R.; Davies-Hill, Theresa M.; Koehler-Stec, Ellen M.; Vannucci, Susan J.; Horne, McDonald K.; Simpson, Ian A.

In: Biochemical Journal, Vol. 328, No. 2, 01.12.1997, p. 511-516.

Research output: Contribution to journalArticle

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Sorbara LR, Davies-Hill TM, Koehler-Stec EM, Vannucci SJ, Horne MK, Simpson IA. Thrombin-induced translocation of GLUT3 glucose transporters in human platelets. Biochemical Journal. 1997 Dec 1;328(2):511-516. https://doi.org/10.1042/bj3280511