Regulation of Glycogen Synthase by Glucose and Glycogen
A Possible Role for AMP-Activated Protein Kinase
- 1School of Biochemistry and Genetics, Medical School, University of Newcastle, Newcastle upon Tyne, U.K.
- 2Cellular Stress Group, MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, London, U.K.
- 3Target Cell Biology, Novo Nordisk A/S, Bagsvaerd, Denmark
We report here use of human myoblasts in culture to study the relationships between cellular glycogen concentrations and the activities of glycogen synthase (GS) and AMP-activated protein kinase (AMPK). Incubation of cells for 2 h in the absence of glucose led to a 25% decrease in glycogen content and a significant decrease in the fractional activity of GS. This was accompanied by stimulation of both the α1 and α2 isoforms of AMPK, without significant alterations in the ratios of adenine nucleotides. When glucose was added to glycogen-depleted cells, a rapid and substantial increase in GS activity was accompanied by inactivation of AMPK back to basal values. Inclusion of the glycogen phosphorylase inhibitor, CP-91149, prevented the loss of glycogen during glucose deprivation but not the activation of AMPK. However, in the absence of prior glycogen breakdown, glucose treatment failed to activate GS above control values, indicating the crucial role of glycogen content. Activation of AMPK by either 5-aminoimidazole-4-carboxamide 1-β-d-ribofuranoside (AICAR) or hydrogen peroxide was also associated with a decrease in the activity ratio of GS. AICAR treatment had no effect on total cellular glycogen content but led to a modest increase in glucose uptake. These data support a role for AMPK in both stimulating glucose uptake and inhibiting GS in intact cells, thus promoting glucose flux through glycolysis.
Address correspondence and reprint requests to Stephen J. Yeaman, School of Biochemistry and Genetics, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, U.K. E-mail:.
Received for publication 28 March 2001 and accepted in revised form 24 September 2002.
J.G.M. is currently affiliated with OSI Pharmaceuticals Ltd., Oxford, U.K.
R.H. receives consulting fees from Xcellsyz, Ltd., a start-up company engaged in deriving immortalized cell lines for study of diabetes. J.G.M. is employed by and holds stock in Novo Nordisk A/S. D.C. is on the Scientific Advisory Board for Xcellsyz. S.J.Y. holds stock in Xcellsyz and has received honoraria from Novo Nordisk and Glaxo Wellcome.
AICAR, 5-aminoimidazole-4-carboxamide 1-β-d-ribofuranoside; AMPK, AMP-activated protein kinase; DME Glu−, glucose-free Dulbecco’s Modified Eagle’s medium; DTT, diothiothreitol; GS, glycogen synthase; GSK3, GS kinase 3; PKA, cAMP-dependent protein kinase; PMSF, phenylmethylsulfonyl fluoride.