Kinetic evidence for unique regulation of GLUT4 trafficking by insulin and AMP-activated protein kinase activators in L6 myotubes

D J Fazakerley, G D Holman, A Marley, D E James, J Stockli, A C F Coster

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Abstract

In L6 myotubes, redistribution of a hemagglutinin (HA) epitope-tagged GLUT4 (HA-GLUT4) to the cell surface occurs rapidly in response to insulin stimulation and AMP-activated protein kinase (AMPK) activation. We have examined whether these separate signaling pathways have a convergent mechanism that leads to GLUT4 mobilization and to changes in GLUT4 recycling. HA antibody uptake on GLUT4 in the basal steady state reached a final equilibrium level that was only 81% of the insulin-stimulated level. AMPK activators (5-aminoimidazole-4-carboxyamide ribonucleoside (AICAR) and A-769662) led to a similar level of antibody uptake to that found in insulin-stimulated cells. However, the combined responses to insulin stimulation and AMPK activation led to an antibody uptake level of similar to 20% above the insulin level. Increases in antibody uptake due to insulin, but not AICAR or A-769662, treatment were reduced by both wortmannin and Akt inhibitor. The GLUT4 internalization rate constant in the basal steady state was very rapid (0.43 min(-1)) and was decreased during the steady-state responses to insulin (0.18 min(-1)), AICAR (0.16 min(-1)), and A-769662 (0.24 min(-1)). This study has revealed a nonconvergent mobilization of GLUT4 in response to activation of Akt and AMPK signaling. Furthermore, GLUT4 trafficking in L6 muscle cells is very reliant on regulated endocytosis for control of cell surface GLUT4 levels.
Original languageEnglish
Pages (from-to)1653-1660
Number of pages8
JournalJournal of Biological Chemistry
Volume285
Issue number3
DOIs
Publication statusPublished - 15 Jan 2010

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AMP-Activated Protein Kinases
Skeletal Muscle Fibers
A 769662
Insulin
Kinetics
Ribonucleosides
Antibodies
Chemical activation
Hemagglutinins
Recycling
Endocytosis
Muscle Cells
Muscle
Epitopes
Rate constants
Cells
4-aminoimidazole

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Kinetic evidence for unique regulation of GLUT4 trafficking by insulin and AMP-activated protein kinase activators in L6 myotubes. / Fazakerley, D J; Holman, G D; Marley, A; James, D E; Stockli, J; Coster, A C F.

In: Journal of Biological Chemistry, Vol. 285, No. 3, 15.01.2010, p. 1653-1660.

Research output: Contribution to journalArticle

Fazakerley, D J ; Holman, G D ; Marley, A ; James, D E ; Stockli, J ; Coster, A C F. / Kinetic evidence for unique regulation of GLUT4 trafficking by insulin and AMP-activated protein kinase activators in L6 myotubes. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 3. pp. 1653-1660.
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