SIRT1, AMP-activated protein kinase phosphorylation and downstream kinases in response to a single bout of sprint exercise: influence of glucose ingestion


This study was designed to examine potential in vivo mechanisms of AMP-activated protein kinase (AMPK) phosphorylation inhibition and its downstream signaling consequences during the recovery period after a single bout of sprint exercise. Sprint exercise induces Thr172-AMPK phosphorylation and increased PGC-1α mRNA, by an unknown mechanism. Muscle biopsies were obtained in 15 young healthy men in response to a 30-s sprint exercise (Wingate test) randomly distributed into two groups: the fasting (n = 7, C) and the glucose group (n = 8, G), who ingested 75 g of glucose 1 h before exercising to inhibit AMPKα phosphorylation. Exercise elicited different patterns of Ser221-ACCβ, Ser473-Akt and Thr642-AS160 phosphorylation, during the recovery period after glucose ingestion. Thirty minutes after the control sprint, Ser485-AMPKα1/Ser491-AMPKα2 phosphorylation was reduced by 33% coinciding with increased Thr172-AMPKα phosphorylation (both, P < 0.05). Glucose abolished the 30-min Thr172-AMPKα phosphorylation. Ser221-ACCβ phosphorylation was elevated immediately following and 30 min after exercise in C and G, implying a dissociation between Thr172-AMPKα and Ser221-ACCβ phosphorylation. Two hours after the sprint, PGC-1α protein expression remained unchanged while SIRT1 (its upstream deacetylase) was increased. Glucose ingestion abolished the SIRT1 response without any significant effect on PGC-1α protein expression. In conclusion, glucose ingestion prior to a sprint exercise profoundly affects Thr172-AMPKα phosphorylation and its downstream signaling during the recovery period.

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  1. Abbot EL, McCormack JG, Reynet C, Hassall DG, Buchan KW, Yeaman SJ (2005) Diverging regulation of pyruvate dehydrogenase kinase isoform gene expression in cultured human muscle cells. FEBS J 272:3004–3014

  2. Ara I, Vicente-Rodriguez G, Jimenez-Ramirez J, Dorado C, Serrano-Sanchez JA, Calbet JA (2004) Regular participation in sports is associated with enhanced physical fitness and lower fat mass in prepubertal boys. Int J Obes Relat Metab Disord 28:1585–1593

  3. Aronson D, Wojtaszewski JF, Thorell A, Nygren J, Zangen D, Richter EA, Ljungqvist O, Fielding RA, Goodyear LJ (1998) Extracellular-regulated protein kinase cascades are activated in response to injury in human skeletal muscle. Am J Physiol 275:C555–C561

  4. Bandyopadhyay GK, Yu JG, Ofrecio J, Olefsky JM (2006) Increased malonyl-CoA levels in muscle from obese and type 2 diabetic subjects lead to decreased fatty acid oxidation and increased lipogenesis; thiazolidinedione treatment reverses these defects. Diabetes 55:2277–2285

  5. Beauloye C, Marsin AS, Bertrand L, Krause U, Hardie DG, Vanoverschelde JL, Hue L (2001) Insulin antagonizes AMP-activated protein kinase activation by ischemia or anoxia in rat hearts, without affecting total adenine nucleotides. FEBS Lett 505:348–352

  6. Birk JB, Wojtaszewski JF (2006) Predominant alpha2/beta2/gamma3 AMPK activation during exercise in human skeletal muscle. J Physiol 577:1021–1032

  7. Calbet JA, Chavarren J, Dorado C (1997) Fractional use of anaerobic capacity during a 30- and a 45-s Wingate test. Eur J Appl Physiol 76:308–313

  8. Calbet JA, De Paz JA, Garatachea N, Cabeza de Vaca S, Chavarren J (2003) Anaerobic energy provision does not limit Wingate exercise performance in endurance-trained cyclists. J Appl Physiol 94:668–676

  9. Canto C, Gerhart-Hines Z, Feige JN, Lagouge M, Noriega L, Milne JC, Elliott PJ, Puigserver P, Auwerx J (2009) AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity. Nature 458:1056–1060

  10. Carling D (2004) The AMP-activated protein kinase cascade—a unifying system for energy control. Trends Biochem Sci 29:18–24

  11. Chabi B, Adhihetty PJ, O’Leary MF, Menzies KJ, Hood DA (2009) Relationship between Sirt1 expression and mitochondrial proteins during conditions of chronic muscle use and disuse. J Appl Physiol 107:1730–1735

  12. Chen ZP, McConell GK, Michell BJ, Snow RJ, Canny BJ, Kemp BE (2000) AMPK signaling in contracting human skeletal muscle: acetyl-CoA carboxylase and NO synthase phosphorylation. Am J Physiol Endocrinol Metab 279:E1202–E1206

  13. Chen ZP, Stephens TJ, Murthy S, Canny BJ, Hargreaves M, Witters LA, Kemp BE, McConell GK (2003) Effect of exercise intensity on skeletal muscle AMPK signaling in humans. Diabetes 52:2205–2212

  14. Christ-Roberts CY, Pratipanawatr T, Pratipanawatr W, Berria R, Belfort R, Mandarino LJ (2003) Increased insulin receptor signaling and glycogen synthase activity contribute to the synergistic effect of exercise on insulin action. J Appl Physiol 95:2519–2529

  15. Davies SP, Hawley SA, Woods A, Carling D, Haystead TA, Hardie DG (1994) Purification of the AMP-activated protein kinase on ATP-gamma-sepharose and analysis of its subunit structure. Eur J Biochem 223:351–357

  16. Davies SP, Helps NR, Cohen PT, Hardie DG (1995) 5′-AMP inhibits dephosphorylation, as well as promoting phosphorylation, of the AMP-activated protein kinase. Studies using bacterially expressed human protein phosphatase-2C alpha and native bovine protein phosphatase-2AC. FEBS Lett 377:421–425

  17. De Bock K, Derave W, Eijnde BO, Hesselink MK, Koninckx E, Rose AJ, Schrauwen P, Bonen A, Richter EA, Hespel P (2008) Effect of training in the fasted state on metabolic responses during exercise with carbohydrate intake. J Appl Physiol 104:1045–1055

  18. Deldicque L, De Bock K, Maris M, Ramaekers M, Nielens H, Francaux M, Hespel P (2009) Increased p70(s6k) phosphorylation during intake of a protein-carbohydrate drink following resistance exercise in the fasted state. Eur J Appl Physiol. doi:10.1007/s00421-009-1289-x

  19. Drummond MJ, Fry CS, Glynn EL, Dreyer HC, Dhanani S, Timmerman KL, Volpi E, Rasmussen BB (2009) Rapamycin administration in humans blocks the contraction-induced increase in skeletal muscle protein synthesis. J Physiol 587:1535–1546

  20. Dzamko N, Schertzer JD, Ryall JG, Steel R, Macaulay SL, Wee S, Chen ZP, Michell BJ, Oakhill JS, Watt MJ, Jorgensen SB, Lynch GS, Kemp BE, Steinberg GR (2008) AMPK-independent pathways regulate skeletal muscle fatty acid oxidation. J Physiol 586:5819–5831

  21. Fujii N, Hayashi T, Hirshman MF, Smith JT, Habinowski SA, Kaijser L, Mu J, Ljungqvist O, Birnbaum MJ, Witters LA, Thorell A, Goodyear LJ (2000) Exercise induces isoform-specific increase in 5′AMP-activated protein kinase activity in human skeletal muscle. Biochem Biophys Res Commun 273:1150–1155

  22. Gamble J, Lopaschuk GD (1997) Insulin inhibition of 5′ adenosine monophosphate-activated protein kinase in the heart results in activation of acetyl coenzyme A carboxylase and inhibition of fatty acid oxidation. Metabolism 46:1270–1274

  23. Gerhart-Hines Z, Rodgers JT, Bare O, Lerin C, Kim SH, Mostoslavsky R, Alt FW, Wu Z, Puigserver P (2007) Metabolic control of muscle mitochondrial function and fatty acid oxidation through SIRT1/PGC-1alpha. EMBO J 26:1913–1923

  24. Gibala MJ, McGee SL, Garnham AP, Howlett KF, Snow RJ, Hargreaves M (2009) Brief intense interval exercise activates AMPK and p38 MAPK signaling and increases the expression of PGC-1alpha in human skeletal muscle. J Appl Physiol 106:929–934

  25. Guerra B, Santana A, Fuentes T, Delgado-Guerra S, Cabrera-Socorro A, Dorado C, Calbet JA (2007) Leptin receptors in human skeletal muscle. J Appl Physiol 102:1786–1792

  26. Gurd BJ, Yoshida Y, Lally J, Holloway GP, Bonen A (2009) The deacetylase enzyme SIRT1 is not associated with oxidative capacity in rat heart and skeletal muscle and its overexpression reduces mitochondrial biogenesis. J Physiol 587:1817–1828

  27. Hardie DG, Carling D, Carlson M (1998) The AMP-activated/SNF1 protein kinase subfamily: metabolic sensors of the eukaryotic cell? Annu Rev Biochem 67:821–855

  28. Hardie DG, Scott JW, Pan DA, Hudson ER (2003) Management of cellular energy by the AMP-activated protein kinase system. FEBS Lett 546:113–120

  29. Hardie DG, Hawley SA, Scott JW (2006) AMP-activated protein kinase—development of the energy sensor concept. J Physiol 574:7–15

  30. Hawley SA, Boudeau J, Reid JL, Mustard KJ, Udd L, Makela TP, Alessi DR, Hardie DG (2003) Complexes between the LKB1 tumor suppressor, STRAD alpha/beta and MO25 alpha/beta are upstream kinases in the AMP-activated protein kinase cascade. J Biol 2:28

  31. Hojlund K, Mustard KJ, Staehr P, Hardie DG, Beck-Nielsen H, Richter EA, Wojtaszewski JF (2004) AMPK activity and isoform protein expression are similar in muscle of obese subjects with and without type 2 diabetes. Am J Physiol Endocrinol Metab 286:E239–E244

  32. Horman S, Vertommen D, Heath R, Neumann D, Mouton V, Woods A, Schlattner U, Wallimann T, Carling D, Hue L, Rider MH (2006) Insulin antagonizes ischemia-induced Thr172 phosphorylation of AMP-activated protein kinase alpha-subunits in heart via hierarchical phosphorylation of Ser485/491. J Biol Chem 281:5335–5340

  33. Hou X, Xu S, Maitland-Toolan KA, Sato K, Jiang B, Ido Y, Lan F, Walsh K, Wierzbicki M, Verbeuren TJ, Cohen RA, Zang M (2008) SIRT1 regulates hepatocyte lipid metabolism through activating AMP-activated protein kinase. J Biol Chem 283:20015–20026

  34. Howlett KF, Mathews A, Garnham A, Sakamoto K (2008) The effect of exercise and insulin on AS160 phosphorylation and 14–3-3 binding capacity in human skeletal muscle. Am J Physiol Endocrinol Metab 294:E401–E407

  35. Jorgensen SB, Viollet B, Andreelli F, Frosig C, Birk JB, Schjerling P, Vaulont S, Richter EA, Wojtaszewski JF (2004) Knockout of the alpha2 but not alpha1 5′-AMP-activated protein kinase isoform abolishes 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside but not contraction-induced glucose uptake in skeletal muscle. J Biol Chem 279:1070–1079

  36. Kovacic S, Soltys CL, Barr AJ, Shiojima I, Walsh K, Dyck JR (2003) Akt activity negatively regulates phosphorylation of AMP-activated protein kinase in the heart. J Biol Chem 278:39422–39427

  37. Leick L, Wojtaszewski JF, Johansen ST, Kiilerich K, Comes G, Hellsten Y, Hidalgo J, Pilegaard H (2008) PGC-1alpha is not mandatory for exercise- and training-induced adaptive gene responses in mouse skeletal muscle. Am J Physiol Endocrinol Metab 294:E463–E474

  38. Long YC, Zierath JR (2008) Influence of AMP-activated protein kinase and calcineurin on metabolic networks in skeletal muscle. Am J Physiol Endocrinol Metab 295:E545–E552

  39. Milner J (2009) Cellular regulation of SIRT1. Curr Pharm Des 15:39–44

  40. Norman B, Sabina RL, Jansson E (2001) Regulation of skeletal muscle ATP catabolism by AMPD1 genotype during sprint exercise in asymptomatic subjects. J Appl Physiol 91:258–264

  41. Perez-Gomez J, Olmedillas H, Delgado-Guerra S, Royo IA, Vicente-Rodriguez G, Ortiz RA, Chavarren J, Calbet JA (2008a) Effects of weight lifting training combined with plyometric exercises on physical fitness, body composition, and knee extension velocity during kicking in football. Appl Physiol Nutr Metab 33:501–510

  42. Perez-Gomez J, Rodriguez GV, Ara I, Olmedillas H, Chavarren J, Gonzalez-Henriquez JJ, Dorado C, Calbet JA (2008b) Role of muscle mass on sprint performance: gender differences? Eur J Appl Physiol 102:685–694

  43. Puigserver P, Spiegelman BM (2003) Peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1 alpha): transcriptional coactivator and metabolic regulator. Endocr Rev 24:78–90

  44. Putman CT, Jones NL, Lands LC, Bragg TM, Hollidge-Horvat MG, Heigenhauser GJ (1995) Skeletal muscle pyruvate dehydrogenase activity during maximal exercise in humans. Am J Physiol 269:E458–E468

  45. Roepstorff C, Thiele M, Hillig T, Pilegaard H, Richter EA, Wojtaszewski JF, Kiens B (2006) Higher skeletal muscle alpha2AMPK activation and lower energy charge and fat oxidation in men than in women during submaximal exercise. J Physiol 574:125–138

  46. Sahlin K, Soderlund K, Tonkonogi M, Hirakoba K (1997) Phosphocreatine content in single fibers of human muscle after sustained submaximal exercise. Am J Physiol 273:C172–C178

  47. Sakamoto K, McCarthy A, Smith D, Green KA, Grahame Hardie D, Ashworth A, Alessi DR (2005) Deficiency of LKB1 in skeletal muscle prevents AMPK activation and glucose uptake during contraction. EMBO J 24:1810–1820

  48. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

  49. Soltys CL, Kovacic S, Dyck JR (2006) Activation of cardiac AMP-activated protein kinase by LKB1 expression or chemical hypoxia is blunted by increased Akt activity. Am J Physiol Heart Circ Physiol 290:H2472–H2479

  50. Sriwijitkamol A, Coletta DK, Wajcberg E, Balbontin GB, Reyna SM, Barrientes J, Eagan PA, Jenkinson CP, Cersosimo E, DeFronzo RA, Sakamoto K, Musi N (2007) Effect of acute exercise on AMPK signaling in skeletal muscle of subjects with type 2 diabetes: a time-course and dose-response study. Diabetes 56:836–848

  51. Suchankova G, Nelson LE, Gerhart-Hines Z, Kelly M, Gauthier MS, Saha AK, Ido Y, Puigserver P, Ruderman NB (2009) Concurrent regulation of AMP-activated protein kinase and SIRT1 in mammalian cells. Biochem Biophys Res Commun 378:836–841

  52. Suwa M, Nakano H, Radak Z, Kumagai S (2008) Endurance exercise increases the SIRT1 and peroxisome proliferator-activated receptor gamma coactivator-1alpha protein expressions in rat skeletal muscle. Metabolism 57:986–998

  53. Turinsky J, Damrau-Abney A (1999) Akt kinases and 2-deoxyglucose uptake in rat skeletal muscles in vivo: study with insulin and exercise. Am J Physiol 276:R277–R282

  54. Walker KS, Deak M, Paterson A, Hudson K, Cohen P, Alessi DR (1998) Activation of protein kinase B beta and gamma isoforms by insulin in vivo and by 3-phosphoinositide-dependent protein kinase-1 in vitro: comparison with protein kinase B alpha. Biochem J 331(Pt 1):299–308

  55. Wilkinson SB, Phillips SM, Atherton PJ, Patel R, Yarasheski KE, Tarnopolsky MA, Rennie MJ (2008) Differential effects of resistance and endurance exercise in the fed state on signalling molecule phosphorylation and protein synthesis in human muscle. J Physiol 586:3701–3717

  56. Witters LA, Kemp BE (1992) Insulin activation of acetyl-CoA carboxylase accompanied by inhibition of the 5′-AMP-activated protein kinase. J Biol Chem 267:2864–2867

  57. Wojtaszewski JF, Nielsen P, Hansen BF, Richter EA, Kiens B (2000) Isoform-specific and exercise intensity-dependent activation of 5′-AMP-activated protein kinase in human skeletal muscle. J Physiol 528(Pt 1):221–226

  58. Wojtaszewski JF, Mourtzakis M, Hillig T, Saltin B, Pilegaard H (2002) Dissociation of AMPK activity and ACCbeta phosphorylation in human muscle during prolonged exercise. Biochem Biophys Res Commun 298:309–316

  59. Woods A, Johnstone SR, Dickerson K, Leiper FC, Fryer LG, Neumann D, Schlattner U, Wallimann T, Carlson M, Carling D (2003a) LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. Curr Biol 13:2004–2008

  60. Woods A, Vertommen D, Neumann D, Turk R, Bayliss J, Schlattner U, Wallimann T, Carling D, Rider MH (2003b) Identification of phosphorylation sites in AMP-activated protein kinase (AMPK) for upstream AMPK kinases and study of their roles by site-directed mutagenesis. J Biol Chem 278:28434–28442

  61. Yeo WK, Lessard SJ, Chen ZP, Garnham AP, Burke LM, Rivas DA, Kemp BE, Hawley JA (2008) Fat adaptation followed by carbohydrate restoration increases AMPK activity in skeletal muscle from trained humans. J Appl Physiol 105:1519–1526

  62. Yu M, Stepto NK, Chibalin AV, Fryer LG, Carling D, Krook A, Hawley JA, Zierath JR (2003) Metabolic and mitogenic signal transduction in human skeletal muscle after intense cycling exercise. J Physiol 546:327–335

  63. Zschoernig B, Mahlknecht U (2008) SIRTUIN 1: regulating the regulator. Biochem Biophys Res Commun 376:251–255

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This study was supported by grants from the Ministerio de Educación y Ciencia (BFI2003-09638, BFU2006-13784 and FEDER) and the Gobierno de Canarias (PI2005/177) and FUNCIS (PI/10/07). Borja Guerra is a fellow of the “Recursos Humanos y Difusión de la Investigación” Programe (ISCIII, MSC, Spain). Special thanks are given to José Navarro de Tuero and Oscar Bernales for his excellent technical assistance. The specialized advice from Tony Webster in editing the English version of the manuscript is also acknowledged. Special thanks are given to all subjects who volunteered for these experiments.

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Correspondence to José A. L. Calbet.

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Communicated by Susan Ward.

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Guerra, B., Guadalupe-Grau, A., Fuentes, T. et al. SIRT1, AMP-activated protein kinase phosphorylation and downstream kinases in response to a single bout of sprint exercise: influence of glucose ingestion. Eur J Appl Physiol 109, 731–743 (2010).

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  • Signaling
  • Wingate
  • Human
  • Skeletal muscle
  • Fatigue
  • Performance