Molecular and Cellular Biochemistry

, Volume 366, Issue 1–2, pp 31–39 | Cite as

Characterization of the high-affinity uptake of fructose-1,6-bisphosphate by cardiac myocytes

  • Thomas J. Wheeler
  • Sufan Chien


Previously, we reported that fructose-1,6-bisphosphate (FBP) was taken up by rat cardiac myocytes by two processes: a component that was saturable at micromolar levels and a nonsaturable component that dominated at millimolar levels. Here, we continued to characterize the saturable high-affinity component, with the aim of identifying the physiological substrate and role for this activity. ATP, ADP, and AMP inhibited the uptake of FBP with apparent affinities of 0.2–0.5 mM. Fumarate and succinate were very weak inhibitors. Several phosphorylated sugars (ribulose-1,5-phosphate, fructose-1-phosphate, ribose-5-phosphate, and inositol-2-phosphate) inhibited FBP uptake with apparent affinities of 40–500 μM. As in our previous study, no tested compound appeared to bind as well as FBP. The data suggest that the best ligands have two phosphoryl groups separated by at least 8 Å. The rates of FBP uptake were measured from 3° to 37°. The calculated activation energy was 15–50 kJ/mol, similar to other membrane transport processes. Uptake of FBP was tested in several types of cells other than cardiac myocytes, and compared to the uptake of 2-deoxyglucose and l-glucose. While FBP uptake in excess of that of l-glucose was observed in some cells, in no case was the uptake as high as in cardiac myocytes. The physiological substrate and role for the high-affinity FBP uptake activity remain unknown.


Cardiac myocytes Fructose-1,6-bisphosphate Transport Adenine nucleotides Dicarboxylate Activation energy 



We thank the following colleagues for providing cells: Vilius Stribinskis, Ph.D., and Kenneth Ramos, Ph.D., mouse aortic smooth muscle cells; Barbara Clark, Ph.D., MA-10 cells and Y-1 cells; Sadhak Sengupta, Ph.D., and Thomas Mitchell, Ph.D., mouse splenocytes; Carolyn Klinge, Ph.D., A549, MCF-7, and human umbilical vein epithelial cells; and Stephanie Webb, Ph.D., and Russell Prough, Ph.D., mouse hepatocytes. Preliminary experiments related to measuring uptake in various cells were performed by Dana Ho, who was supported by the Kentucky Biomedical Research Infrastructures Network undergraduate summer research program. We also thank Mary Anne Hauck for technical assistance and Robert D. Gray, Ph.D., for advice concerning molecular structures. This study was supported in part by National Institutes of Health grant HL64186.


  1. 1.
    Hirokawa F, Nakai T, Yamaue H (2002) Storage solution containing fructose-1,6-bisphosphate inhibits the excess activation of Kupffer cells in cold liver preservation. Transplantation 74:779–783PubMedCrossRefGoogle Scholar
  2. 2.
    Didlake R, Kirchner KA, Lewin J, Bower JD, Markov A (1985) Protection from ischemic renal injury by fructose-1,6-diphosphate infusion in the rat. Circ Shock 16:205–2512PubMedGoogle Scholar
  3. 3.
    Bickler PE, Buck LT (1996) Effects of fructose-1,6-bisphosphate on glutamate release and ATP loss from rat brain slices during hypoxia. J Neurochem 67:1463–1468PubMedCrossRefGoogle Scholar
  4. 4.
    Hardin CD, Roberts TM (1994) Metabolism of exogenously applied fructose 1,6-bisphosphate in hypoxic vascular smooth muscle. Am J Physiol 267:H2325–H2332PubMedGoogle Scholar
  5. 5.
    Chu SJ, Chang DM, Wang D, Chen YH, Hsu CW, Hsu K (2002) Fructose-1,6-diphosphate attenuates acute lung injury induced by ischemia-reperfusion in rats. Crit Care Med 30:1605–1609PubMedCrossRefGoogle Scholar
  6. 6.
    Sun JX, Farias LA, Markov AK (1990) Fructose 1-6 diphosphate prevents intestinal ischemic reperfusion injury and death in rats. Gastroenterology 98:117–126PubMedGoogle Scholar
  7. 7.
    Niu W, Zhang F, Ehringer W, Tseng M, Gray L, Chien S (1999) Enhancement of hypothermic heart preservation with fructose 1,6-diphosphate. J Surg Res 85:120–129PubMedCrossRefGoogle Scholar
  8. 8.
    Chien S, Zhang F, Niu W, Ehringer W, Chiang B, Shi X, Gray LA (2000) Fructose-1,6-diphosphate and a glucose-free solution enhances functional recovery in hypothermic heart preservation. J Heart Lung Transpl 19:277–285CrossRefGoogle Scholar
  9. 9.
    Hua D, Zhuang X, Ye J, Wilson D, Chiang B, Chien S (2003) Using fructose-1,6-diphosphate during hypothermic rabbit-heart preservation: a high-energy phosphate study. J Heart Lung Transpl 22:574–582CrossRefGoogle Scholar
  10. 10.
    Wheeler TJ, Wiegand CB, Chien S (2005) Fructose-1,6-bisphosphate enhances hypothermic preservation of cardiac myocytes. J Heart Lung Transpl 24:1378–1384CrossRefGoogle Scholar
  11. 11.
    Hardin CD, Lazzarino G, Tavazzi B, Di Pierro D, Roberts TM, Giardina B, Rovetto MJ (2001) Myocardial metabolism of exogenous FDP is consistent with transport by a dicarboxylate transporter. Am J Physiol 281:H2654–H2660Google Scholar
  12. 12.
    Lazzarino G, Cattani L, Costrini R, Mulieri L, Candiani A, Galzigna L (1984) Increase of intraerythrocytic fructose-1,6-diphosphate after incubation of whole human blood with fructose-1,6-diphosphate. Clin Biochem 17:42–45PubMedCrossRefGoogle Scholar
  13. 13.
    Gregory GA, Yu ACH, Chan PH (1989) Fructose-1,6-diphosphate protects astrocytes from hypoxic damage. J Cereb Blood Flow Metab 9:29–34PubMedCrossRefGoogle Scholar
  14. 14.
    Kelleher JA, Chan PH, Chan TYY, Gregory GA (1995) Energy metabolism in hypoxic astrocytes: protective mechanisms of fructose-1,6-bisphosphate. Neurochem Res 20:785–792PubMedCrossRefGoogle Scholar
  15. 15.
    Juergens TM, Hardin CD (1996) Fructose-1,6-bisphosphate as a metabolic substrate in hog ileum smooth muscle during hypoxia. Mol Cell Biochem 154:83–93PubMedCrossRefGoogle Scholar
  16. 16.
    Tavazzi B, Starnes JW, Lazzarino G, DiPierro D, Nuutinen EM, Giardina B (1992) Exogenous fructose-1,6-bisphosphate is a metabolizable substrate for the isolated normoxic rat heart. Basic Res Cardiol 87:280–289PubMedCrossRefGoogle Scholar
  17. 17.
    Takeuchi K, Cao-Danh H, Friehs I, Glynn P, D’Agostino D, Simplaceanu E, McGowan FX, del Nido PJ (1998) Administration of fructose 1,6-diphosphate during early reperfusion significantly improves recovery of contractile function in the postischemic heart. J Thorac Cardiovasc Surg 116:335–343PubMedCrossRefGoogle Scholar
  18. 18.
    Ehringer WD, Niue W, Chiang B, Wang OL, Gordon L, Chien S (2000) Membrane permeability of fructose-1,6-diphosphate in lipid vesicles and endothelial cells. Mol Cell Biochem 210:23–45CrossRefGoogle Scholar
  19. 19.
    Ehringer WD, Chiang B, Chien S (2001) The uptake and metabolism of fructose-1,6-diphosphate in rat cardiomyocytes. Mol Cell Biochem 221:33–40PubMedCrossRefGoogle Scholar
  20. 20.
    Wheeler TJ, McCurdy JM, denDekker A, Chien S (2004) Permeability of fructose-1,6-bisphosphate in liposomes and cardiac myocytes. Mol Cell Biochem 259:105–114PubMedCrossRefGoogle Scholar
  21. 21.
    Vassort G (2001) Adenosine 5′-triphosphate: a P2-purinergic agonist in the myocardium. Physiol Rev 81:767–806PubMedGoogle Scholar
  22. 22.
    Colston VL, Wheeler TJ (2001) Stimulation of cardiac glucose transport by inhibitors of oxidative phosphorylation. Life Sci 69:2383–2398PubMedCrossRefGoogle Scholar
  23. 23.
    Fischer Y, Rose H, Kammermeier K (1991) Highly insulin-responsive isolated rat heart muscle cells yielded by a modified isolation method. Life Sci 49:1679–1688PubMedCrossRefGoogle Scholar
  24. 24.
    Mellor KM, Ritchie RH, Davidoff AJ, Delbridge LMD (2010) Elevated dietary sugar and the heart: experimental models and myocardial remodeling. Can J Physiol Pharmacol 88:525–540PubMedCrossRefGoogle Scholar
  25. 25.
    Galzigna L, Rizzoli V, Bianchi M, Rigobello MP, Scuri R (1989) Some effects of fructose-1,6-diphosphate on rat myocardial tissue related to a membrane-stabilizing action. Cell Biochem Function 7:91–96CrossRefGoogle Scholar
  26. 26.
    Hu XJ, Peng F, Zhou HQ, Zhang ZH, Cheng WY, Feng HF (2000) The abnormality of glucose transporter in the erythrocyte membrane of Chinese type 2 diabetic patients. Biochim Biophys Acta 1466:306–314PubMedCrossRefGoogle Scholar
  27. 27.
    Zeidel ML, Ambudkar SV, Smith BL, Agre P (1992) Reconstitution of functional water channels in liposomes containing purified red cell CHIP28 protein. Biochemistry 31:7436–7440PubMedCrossRefGoogle Scholar
  28. 28.
    Richards OC, Rutter WJ (1961) Comparative properties of yeast and muscle aldolase. J Biol Chem 236:3185–319227PubMedGoogle Scholar
  29. 29.
    Wheeler TJ (1986) Kinetics of glucose transport in human erythrocytes: zero-trans efflux and infinite-trans efflux at 0 °C. Biochim Biophys Acta 862:387–398PubMedCrossRefGoogle Scholar
  30. 30.
    Franke L, Schewe HJ, Müller B, Campman V, Kitzrow W, Uebelhack R, Berghöfer A, Müller-Oerlinghausen B (2000) Serotonergic platelet variables in unmedicated patients suffering from major depression and healthy subjects: relationship between 5HT content and 5HT uptake. Life Sci 67:301–315PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular BiologyUniversity of Louisville School of MedicineLouisvilleUSA
  2. 2.Department of SurgeryUniversity of Louisville School of Medicine, Health Sciences CenterLouisvilleUSA

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