Systems and Synthetic Biology

, Volume 1, Issue 1, pp 47–58 | Cite as

The Hsp70 chaperone system maintains high concentrations of active proteins and suppresses ATP consumption during heat shock

Research Article


Hsp70 chaperones assist protein folding by cycling between the ATP-bound T state with low affinity for substrates and the ADP-bound R state with high affinity for substrates. The transition from the T to R state is catalyzed by the synergistic action of the substrate and DnaJ cochaperones. The reverse transition from the R state to the T state is accelerated by the nucleotide exchange factor GrpE. These two processes, T-to-R and R-to-T conversion, are affected differently by temperature change. Here we modeled Hsp70-mediated protein folding under permanent and transient heat shock based on published experimental data. Our simulation results were in agreement with in vitro wild-type Escherichia coli chaperone experimental data at 25°C and reflected R-to-T ratio dynamics in response to temperature effects. Our simulation results suggested that the chaperone system evolved naturally to maintain the concentration of active protein as high as possible during heat shock, even at the cost of recovered activity after return to optimal growth conditions. They also revealed that the chaperone system evolved to suppress ATP consumption at non-optimal high growing temperatures.


GrpE Heat-shock Hsp70 Protein folding Systems biology Temperature 



We thank all the members of the E-CELL project at Keio University for their support. This work was supported in part by grants from The 21st Century COE Program “Understanding and Control of Life’s Function via Systems Biology”; from the Leading Project for Biosimulation, Keio University, on behalf of The Ministry of Education, Culture, Sports, Science, and Technology (MEXT, Japan); and from the CREST–JST project “Development of Modeling/Simulation Environments for Systems Biology” on behalf of the Japan Science and Technology Agency.


  1. Agashe VR, Hartl FU (2000) Roles of molecular chaperones in cytoplasmic protein folding. Semin Cell Dev Biol 11(1):15–25PubMedCrossRefGoogle Scholar
  2. Anfinsen CB (1973) Principles that govern the folding of protein chains. Science 181:223–230PubMedCrossRefGoogle Scholar
  3. Brehmer D, Gassler C et al (2004) Influence of GrpE on DnaK-substrate interactions. J Biol Chem 279(27):27957–27964PubMedCrossRefGoogle Scholar
  4. Bukau B, Deuerling E et al (2000) Getting newly synthesized proteins into shape. Cell 101:119–122PubMedCrossRefGoogle Scholar
  5. Chesnokova LS, Slepenkov SV et al (2003) Deletion of DnaK’s lid strengthens binding to the nucleotide exchange factor, GrpE: a kinetic and thermodynamic analysis. Biochemistry 42(30):9028–9040PubMedCrossRefGoogle Scholar
  6. Dobson CM (1999) Protein misfolding, evolution and disease. TiBS 24:329–332PubMedGoogle Scholar
  7. Dobson CM (2006) Protein aggregation and its consequences for human disease. Protein Pept Lett 13(3):219–227PubMedCrossRefGoogle Scholar
  8. Farr CD, Witt SN (1999) ATP lowers the activation enthalpy barriers to DnaK-peptide complex formation and dissociation. Cell Stress Chaperon 4(2):77–85CrossRefGoogle Scholar
  9. Feldman DE, Frydman J (2000) Protein folding in vivo: the importance of molecular chaperones. Curr Opin Struct Biol 10(1):26–33PubMedCrossRefGoogle Scholar
  10. Gamer J, Multhaup G et al. (1996) A cycle of binding and release of the DnaK, DnaJ and GrpE chaperones regulates activity of the Escherichia coli heat shock transcription factor sigma32. Embo J 15(3):607–617PubMedGoogle Scholar
  11. Gething M-JH, Sambrook JF (1992) Protein folding in the cell. Nature 355:33–45PubMedCrossRefGoogle Scholar
  12. Gillespie DT (1976) A general method for numerically simulating the stochastic time evolution of coupled chemical reactions. J Comput Phys 22(4):403–434CrossRefGoogle Scholar
  13. Gisler SM, Pierpaoli EV et al (1998) Catapult mechanism renders the chaperone action of Hsp70 unidirectional. J Mol Biol 279:833–840PubMedCrossRefGoogle Scholar
  14. Grimshaw JP, Jelesarov I et al (2001) Reversible thermal transition in GrpE, the nucleotide exchange factor of the DnaK heat-shock system. J Biol Chem 276(9):6098–6104PubMedCrossRefGoogle Scholar
  15. Grimshaw JP, Jelesarov I et al (2003) Thermosensor action of GrpE: the DnaK chaperone system at heat shock temperatures. J Biol Chem 278(21):19048–19053PubMedCrossRefGoogle Scholar
  16. Groemping Y., Reinstein J (2001) Folding properties of the nucleotide exchange factor GrpE from Thermus thermophilus: GrpE is a thermosensor that mediates heat shock response. J Mol Biol 314(1):167–178PubMedCrossRefGoogle Scholar
  17. Gross CA (1999). Function and Regulation of the Heat Shock Proteins. Escherichia coli and Salmonella Cellular and Molecular Biology. N. F.C., Adobe and Mira Digital PublishingGoogle Scholar
  18. Hartl FU, Hayer-Hartl M (2002) Molecular chaperones in the cytosol: from nascent chain to folded protein. Science 295(5561):1852–1858PubMedCrossRefGoogle Scholar
  19. Herbst R, Gast K et al (1998) Folding of firefly (Photinus pyralis) luciferase: aggregation and reactivation of unfolding intermediates. Biochemistry 37:6586–6597PubMedCrossRefGoogle Scholar
  20. Herbst R., Schäfer U et al (1997) Equilibrium intermediates in the reversible unfolding of firefly (Photinus pyralis) luciferase. J Biol Chem 272:7099–7105PubMedCrossRefGoogle Scholar
  21. Hu B, Mayer MP et al (2006) Modeling hsp70-mediated protein folding. Biophys J 91(2):496–507PubMedCrossRefGoogle Scholar
  22. Karzai AW, McMacken R (1996) A bipartite signaling mechanism involved in DnaJ-mediated activation of the Escherichia coli DnaK protein. J Biol Chem 271(19):11236–11246PubMedCrossRefGoogle Scholar
  23. Langer T, Lu C et al (1992) Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding. Nature 356(6371):683–689PubMedCrossRefGoogle Scholar
  24. Laufen T, Mayer MP et al (1999) Mechanism of regulation of Hsp70 chaperones by DnaJ co-chaperones. Proc Natl Acad Sci USA 96:5452–5457PubMedCrossRefGoogle Scholar
  25. Liberek K, Marszalek J et al (1991) Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK. Proc Natl Acad Sci USA 88:2874–2878PubMedCrossRefGoogle Scholar
  26. Mayer MP, Brehmer D et al (2001) Hsp70 chaperone machines. Adv Protein Chem 59:1–44PubMedCrossRefGoogle Scholar
  27. Mayer MP, Rudiger S et al (2000a) Molecular basis for interactions of the DnaK chaperone with substrates. Biol Chem 381(9–10):877–885CrossRefGoogle Scholar
  28. Mayer MP, Schröder H et al (2000b) Multistep mechanism of substrate binding determines chaperone activity of Hsp70. Nature Struct Biol 7:586–593CrossRefGoogle Scholar
  29. McCarty JS, Buchberger A et al (1995) The role of ATP in the functional cycle of the DnaK chaperone system. J Mol Biol 249:126–137PubMedCrossRefGoogle Scholar
  30. Mogk A, Tomoyasu T et al (1999) Identification of thermolabile E. coli proteins: prevention and reversion of aggregation by DnaK and ClpB. EMBO J 18:6934–6949PubMedCrossRefGoogle Scholar
  31. Packschies L, Theyssen H et al (1997) GrpE accelerates nucleotide exchange of the molecular chaperone DnaK with an associative displacement mechanism. Biochemistry 36(12):3417–3422PubMedCrossRefGoogle Scholar
  32. Radford SE (2000) Protein folding: progress made and promises ahead. Trends Biochem Sci 25(12):611–618PubMedCrossRefGoogle Scholar
  33. Russell R, Jordan R et al (1998) Kinetic characterization of the ATPase cycle of the DnaK molecular chaperone. Biochemistry 37:596–607PubMedCrossRefGoogle Scholar
  34. Schmid D, Baici A et al (1994) Kinetics of molecular chaperone action. Science 263:971–973PubMedCrossRefGoogle Scholar
  35. Schröder H, Langer T et al (1993) DnaK, DnaJ and GrpE form a cellular chaperone machinery capable of repairing heat-induced protein damage. Embo J 12(11):4137–4144PubMedGoogle Scholar
  36. Takahashi K, Kaizu K et al (2004) A multi-algorithm, multi-timescale method for cell simulation. Bioinformatics 20(4):538–546PubMedCrossRefGoogle Scholar
  37. Theyssen H, Schuster H-P et al (1996) The second step of ATP binding to DnaK induces peptide release. J Mol Biol 263:657–670PubMedCrossRefGoogle Scholar
  38. Tomita M, Hashimoto K et al (1999) E-CELL: software environment for whole-cell simulation. Bioinformatics 15(1):72–84PubMedCrossRefGoogle Scholar
  39. Vogel M, Bukau B et al (2006) Allosteric regulation of Hsp70 chaperones by a proline switch. Mol Cell 21(3):359–367PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science + Business Media B.V. 2007

Authors and Affiliations

  1. 1.Institute for Advanced BiosciencesKeio UniversityTsuruokaJapan
  2. 2.Systems Biology Program, Graduate School of Media and GovernanceKeio University TsuruokaJapan

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