Advertisement

Systems-Level Analysis of Protein Quality in Inclusion Body-Forming Escherichia coli Cells

  • Elena Garcìa-Fruitòs
  • Nuria Gonzàlez-Montalbàn
  • Mònica Martìnez-Alonso
  • Ursula Rinas
  • Antonio Villaverde

Abstract

Recombinant proteins produced in Escherichia coli often aggregate as amorphous masses of insoluble material known as inclusion bodies. Being quite homogeneous in their composition, inclusion bodies display amyloid-like properties such as sequence-dependent protein-protein interactions, seeding-driven deposition of their components and β-sheet intermolecular architecture. However, inclusion bodies formed by different proteins and enzymes also show important extents of native-like secondary structure and include significant proportions of properly folded, functional protein, which makes them suitable to be used in catalytic processes. Inclusion bodies are formed as a result of the incapability of the quality control cell system to cope with the non physiological amounts of misfolding-prone proteins produced upon recombinant gene expression. Multiple cellular proteins involved in the quality control, namely chaperones and proteases, participate in their formation and co-ordinately determine the amount of aggregated protein, the size of aggregates and the main structural and functional properties of the embedded polypeptides, such as their inner molecular organization.

Keywords

Recombinant Protein Inclusion Body Molecular Chaperone Recombinant Protein Production Small Heat Shock Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Allen SP, Polazzi JO, Gierse JK et al. (1992) Two novel heat shock genes encoding proteins produced in response to heterologous protein expression in Escherichia coli. J Bacteriol 174(21):6938–47PubMedGoogle Scholar
  2. Ami D, Bonecchi L, Cali S et al. (2003) FT-IR study of heterologous protein expression in recombinant Escherichia coli strains. Biochim Biophys Acta 1624(1–3):6–10Google Scholar
  3. Ami D, Natalello A, Gatti-Lafranconi P et al. (2005) Kinetics of inclusion body formation studied in intact cells by FT-IR spectroscopy. FEBS Lett 579(16):3433–6PubMedGoogle Scholar
  4. Ami D, Natalello A, Taylor G et al. (2006) Structural analysis of protein inclusion bodies by Fourier transform infrared microspectroscopy. Biochim Biophys Acta 1764(4):793–9PubMedGoogle Scholar
  5. Amrein KE, Takacs B, Stieger M et al. (1995) Purification and characterization of recombinant human p50csk protein-tyrosine kinase from an Escherichia coli expression system overproducing the bacterial chaperones GroES and GroEL. Proc Natl Acad Sci USA 92(4):1048–52PubMedGoogle Scholar
  6. Anfinsen CB (1973) Principles that govern the folding of protein chains. Science 181(96):223–30PubMedGoogle Scholar
  7. Apiyo D, Wittung-Stafshede P (2002) Presence of the cofactor speeds up folding of Desulfovibrio desulfuricans flavodoxin. Protein Sci 11(5):1129–35PubMedGoogle Scholar
  8. Baldwin RL (1986) Temperature dependence of the hydrophobic interaction in protein folding. Proc Natl Acad Sci USA 83(21):8069–72PubMedGoogle Scholar
  9. Baldwin RL (1994) Protein folding. Matching speed and stability. Nature 369(6477):183–4Google Scholar
  10. Baneyx F (2004) Keeping up with protein folding. Microb Cell Fact 3(1):6PubMedGoogle Scholar
  11. Baneyx F, Georgiou G (1991) Construction and characterization of Escherichia coli strains deficient in multiple secreted proteases: protease III degrades high-molecular-weight substrates in vivo. J Bacteriol 173(8):2696–703PubMedGoogle Scholar
  12. Ben-Zvi AP, Goloubinoff P (2001) Review: mechanisms of disaggregation and refolding of stable protein aggregates by molecular chaperones. J Struct Biol 135(2):84–93PubMedGoogle Scholar
  13. Bowden GA, Paredes AM, Georgiou G (1991) Structure and morphology of protein inclusion bodies in Escherichia coli. Biotechnology (N Y) 9(8):725–30Google Scholar
  14. Bruser T, Yano T, Brune DC et al. (2003) Membrane targeting of a folded and cofactor-containing protein. Eur J Biochem 270(6):1211–21PubMedGoogle Scholar
  15. Bucciantini M, Calloni G, Chiti F et al. (2004) Prefibrillar amyloid protein aggregates share common features of cytotoxicity. J Biol Chem 279(30):31374–82PubMedGoogle Scholar
  16. Bucciantini M, Giannoni E, Chiti F et al. (2002) Inherent toxicity of aggregates implies a common mechanism for protein misfolding diseases. Nature 416(6880):507–11PubMedGoogle Scholar
  17. Bukau B (1993) Regulation of the Escherichia coli heat-shock response. Mol Microbiol 9(4): 671–80PubMedGoogle Scholar
  18. Bukau B, Deuerling E, Pfund C et al. (2000) Getting newly synthesized proteins into shape. Cell 101(2):119–22PubMedGoogle Scholar
  19. Bukau B, Weissman J, Horwich A (2006) Molecular chaperones and protein quality control. Cell 125(3):443–51PubMedGoogle Scholar
  20. Carbonell X, Villaverde A (2002) Protein aggregated into bacterial inclusion bodies does not result in protection from proteolytic digestion. Biotechnol Lett 24(23):1939–1944Google Scholar
  21. Carrio M, Gonzalez-Montalban N, Vera A et al. (2005) Amyloid-like properties of bacterial inclusion bodies. J Mol Biol 347(5):1025–37PubMedGoogle Scholar
  22. Carrio MM, Corchero JL, Villaverde A (1998) Dynamics of in vivo protein aggregation: building inclusion bodies in recombinant bacteria. FEMS Microbiol Lett 169(1):9–15PubMedGoogle Scholar
  23. Carrio MM, Corchero JL, Villaverde A (1999) Proteolytic digestion of bacterial inclusion body proteins during dynamic transition between soluble and insoluble forms. Biochim Biophys Acta 1434(1):170–6PubMedGoogle Scholar
  24. Carrio MM, Villaverde A (2001) Protein aggregation as bacterial inclusion bodies is reversible. FEBS Lett 489(1):29–33PubMedGoogle Scholar
  25. Carrio MM, Villaverde A (2002) Construction and deconstruction of bacterial inclusion bodies. J Biotechnol 96(1):3–12PubMedGoogle Scholar
  26. Carrio MM, Villaverde A (2003) Role of molecular chaperones in inclusion body formation. FEBS Lett 537(1–3):215–21PubMedGoogle Scholar
  27. Carrio MM, Villaverde A (2005) Localization of chaperones DnaK and GroEL in bacterial inclusion bodies. J Bacteriol 187(10):3599–601PubMedGoogle Scholar
  28. Chapman D, Haris PI (1989) Biomembrane structures. Fourier transform infrared spectroscopy and biomembrane technology. Biochem Soc Trans 17(6):951–3Google Scholar
  29. Chapman E, Farr GW, Usaite R et al. (2006) Global aggregation of newly translated proteins in an Escherichia coli strain deficient of the chaperonin GroEL. Proc Natl Acad Sci USA 103(43):15800–5PubMedGoogle Scholar
  30. Chesshyre JA, Hipkiss AR (1989) Low temperatures stabilize interferon α-2 against proteolysis in Methylophilus methylotrophus and Escherichia coli. Appl Microbiol Biotechnol 31(2): 158–162Google Scholar
  31. Chiti F, Taddei N, van Nuland NA et al. (1998) Structural characterization of the transition state for folding of muscle acylphosphatase. J Mol Biol 283(4):893–903PubMedGoogle Scholar
  32. Chuang SE, Burland V, Plunkett G, 3rd et al. (1993) Sequence analysis of four new heat-shock genes constituting the hslTS/ibpAB and hslVU operons in Escherichia coli. Gene 134(1):1–6PubMedGoogle Scholar
  33. Clark ED (2001) Protein refolding for industrial processes. Curr Opin Biotechnol 12(2):202–7PubMedGoogle Scholar
  34. Corchero JL, Cubarsi R, Enfors S et al. (1997) Limited in vivo proteolysis of aggregated proteins. Biochem Biophys Res Commun 237(2):325–30PubMedGoogle Scholar
  35. Cubarsi R, Carrio MM, Villaverde A (2005) A mathematical approach to molecular organization and proteolytic disintegration of bacterial inclusion bodies. Math Med Biol 22(3):209–26PubMedGoogle Scholar
  36. Dale GE, Schonfeld HJ, Langen H et al. (1994) Increased solubility of trimethoprim-resistant type S1 DHFR from Staphylococcus aureus in Escherichia coli cells overproducing the chaperonins GroEL and GroES. Protein Eng 7(7):925–31PubMedGoogle Scholar
  37. de Groot NS, Ventura S (2006) Protein activity in bacterial inclusion bodies correlates with predicted aggregation rates. J Biotechnol 125(1):110–3PubMedGoogle Scholar
  38. de Marco A, Deuerling E, Mogk A et al. (2007) Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli. BMC Biotechnol 7:32PubMedGoogle Scholar
  39. de Marco A, Volrath S, Bruyere T et al. (2000) Recombinant maize protoporphyrinogen IX oxidase expressed in Escherichia coli forms complexes with GroEL and DnaK chaperones. Protein Expr Purif 20(1):81–6PubMedGoogle Scholar
  40. Dill KA, Chan HS (1997) From Levinthal to pathways to funnels. Nat Struct Biol 4(1):10–9PubMedGoogle Scholar
  41. Dinner AR, Sali A, Smith LJ et al. (2000) Understanding protein folding via free-energy surfaces from theory and experiment. Trends Biochem Sci 25(7):331–9PubMedGoogle Scholar
  42. Dobson CM (2004) Principles of protein folding, misfolding and aggregation. Semin Cell Dev Biol 15(1):3–16PubMedGoogle Scholar
  43. Echave P, Esparza-Ceron MA, Cabiscol E et al. (2002) DnaK dependence of mutant ethanol oxidoreductases evolved for aerobic function and protective role of the chaperone against protein oxidative damage in Escherichia coli. Proc Natl Acad Sci USA 99(7):4626–31PubMedGoogle Scholar
  44. Ehrnsperger M, Graber S, Gaestel M et al. (1997) Binding of non-native protein to Hsp25 during heat shock creates a reservoir of folding intermediates for reactivation. Embo J 16(2):221–9PubMedGoogle Scholar
  45. Ellis J (1987) Proteins as molecular chaperones. Nature 328(6129):378–9PubMedGoogle Scholar
  46. Ellis RJ (2001) Macromolecular crowding: obvious but underappreciated. Trends Biochem Sci 26(10):597–604PubMedGoogle Scholar
  47. Enfors SO (1992) Control of in vivo proteolysis in the production of recombinant proteins. Trends Biotechnol 10(9):310–5PubMedGoogle Scholar
  48. Esposito D, Chatterjee DK (2006) Enhancement of soluble protein expression through the use of fusion tags. Curr Opin Biotechnol 17(4):353–8PubMedGoogle Scholar
  49. Estapè D, Rinas U (1996) Optimized procedures for purification and solubilization of basic fibroblast growth factor inclusion bodies. Biotechnol. Tech 10(7):481–484Google Scholar
  50. Ewalt KL, Hendrick JP, Houry WA et al. (1997) In vivo observation of polypeptide flux through the bacterial chaperonin system. Cell 90(3):491–500PubMedGoogle Scholar
  51. Fahnert B, Lilie H, Neubauer P (2004) Inclusion bodies: formation and utilisation. Adv Biochem Eng Biotechnol 89:93–142PubMedGoogle Scholar
  52. Fayet O, Ziegelhoffer T, Georgopoulos C (1989) The groES and groEL heat shock gene products of Escherichia coli are essential for bacterial growth at all temperatures. J Bacteriol 171(3): 1379–85PubMedGoogle Scholar
  53. Ferrer M, Chernikova TN, Yakimov MM et al. (2003) Chaperonins govern growth of Escherichia coli at low temperatures. Nat Biotechnol 21(11):1266–7PubMedGoogle Scholar
  54. Fink AL (1998) Protein aggregation: folding aggregates, inclusion bodies and amyloid. Fold Des 3(1):R9–23PubMedGoogle Scholar
  55. Fredriksson A, Ballesteros M, Dukan S et al. (2005) Defense against protein carbonylation by DnaK/DnaJ and proteases of the heat shock regulon. J Bacteriol 187(12):4207–13PubMedGoogle Scholar
  56. Garcia-Fruitos E, Aris A, Villaverde A (2007a) Localization of functional polypeptides in bacterial inclusion bodies. Appl Environ Microbiol 73(1):289–94PubMedGoogle Scholar
  57. Garcia-Fruitos E, Carrio MM, Aris A et al. (2005a) Folding of a misfolding-prone beta-galactosidase in absence of DnaK. Biotechnol Bioeng 90(7):869–75PubMedGoogle Scholar
  58. Garcia-Fruitos E, Gonzalez-Montalban N, Morell M et al. (2005b) Aggregation as bacterial inclusion bodies does not imply inactivation of enzymes and fluorescent proteins. Microb Cell Fact 4:27PubMedGoogle Scholar
  59. Garcia-Fruitos E, Martinez-Alonso M, Gonzalez-Montalban N et al. (2007b) Divergent genetic control of protein solubility and conformational quality in Escherichia coli. J Mol Biol 374(1):195–205PubMedGoogle Scholar
  60. Gasser B, Saloheimo M, Rinas U et al. (2008) Protein folding and conformational stress in microbial cells producing recombinant proteins: a host comparative overview. Microb Cell Fact 7:11PubMedGoogle Scholar
  61. Genevaux P, Georgopoulos C, Kelley WL (2007) The Hsp70 chaperone machines of Escherichia coli: a paradigm for the repartition of chaperone functions. Mol Microbiol 66(4):840–57PubMedGoogle Scholar
  62. Genevaux P, Schwager F, Georgopoulos C et al. (2001) The djlA gene acts synergistically with dnaJ in promoting Escherichia coli growth. J Bacteriol 183(19):5747–50PubMedGoogle Scholar
  63. Georgiou G, Valax P (1996) Expression of correctly folded proteins in Escherichia coli. Curr Opin Biotechnol 7(2):190–7PubMedGoogle Scholar
  64. Georgiou G, Valax P, Ostermeier M et al. (1994) Folding and aggregation of TEM beta-lactamase: analogies with the formation of inclusion bodies in Escherichia coli. Protein Sci 3(11):1953–60PubMedGoogle Scholar
  65. Gething MJ, Sambrook J (1992) Protein folding in the cell. Nature 355(6355):33–45PubMedGoogle Scholar
  66. Glover JR, Lindquist S (1998) Hsp104, Hsp70, and Hsp40: a novel chaperone system that rescues previously aggregated proteins. Cell 94(1):73–82PubMedGoogle Scholar
  67. Glover JR, Tkach JM (2001) Crowbars and ratchets: hsp100 chaperones as tools in reversing protein aggregation. Biochem Cell Biol 79(5):557–68PubMedGoogle Scholar
  68. Goloubinoff P, Mogk A, Zvi AP et al. (1999) Sequential mechanism of solubilization and refolding of stable protein aggregates by a bichaperone network. Proc Natl Acad Sci USA 96(24): 13732–7PubMedGoogle Scholar
  69. Gonzalez-Montalban N, Garcia-Fruitos E, Ventura S et al. (2006) The chaperone DnaK controls the fractioning of functional protein between soluble and insoluble cell fractions in inclusion body-forming cells. Microb Cell Fact 5:26PubMedGoogle Scholar
  70. Gonzalez-Montalban N, Garcia-Fruitos E, Villaverde A (2007a) Recombinant protein solubility – does more mean better? Nat Biotechnol 25(7):718–20Google Scholar
  71. Gonzalez-Montalban N, Natalello A, Garcia-Fruitos E et al. (2008) In situ protein folding and activation in bacterial inclusion bodies. Biotechnol Bioeng 100(4):797–802PubMedGoogle Scholar
  72. Gonzalez-Montalban N, Villaverde A, Aris A (2007b) Amyloid-linked cellular toxicity triggered by bacterial inclusion bodies. Biochem Biophys Res Commun 355(3):637–42PubMedGoogle Scholar
  73. Gottesman S, Wickner S, Maurizi MR (1997) Protein quality control: triage by chaperones and proteases. Genes Dev 11(7):815–23PubMedGoogle Scholar
  74. Graf PC, Jakob U (2002) Redox-regulated molecular chaperones. Cell Mol Life Sci 59(10): 1624–31PubMedGoogle Scholar
  75. Grantcharova V, Alm EJ, Baker D et al. (2001) Mechanisms of protein folding. Curr Opin Struct Biol 11(1):70–82PubMedGoogle Scholar
  76. Grudzielanek S, Velkova A, Shukla A et al. (2007) Cytotoxicity of insulin within its self-assembly and amyloidogenic pathways. J Mol Biol 370(2):372–84PubMedGoogle Scholar
  77. Gupta RS, Singh B (1994) Phylogenetic analysis of 70 kD heat shock protein sequences suggests a chimeric origin for the eukaryotic cell nucleus. Curr Biol 4(12):1104–14PubMedGoogle Scholar
  78. Han MJ, Park SJ, Park TJ et al. (2004) Roles and applications of small heat shock proteins in the production of recombinant proteins in Escherichia coli. Biotechnol Bioeng 88(4): 426–36PubMedGoogle Scholar
  79. Harris TJ, Patel T, Marston FA et al. (1986) Cloning of cDNA coding for human tissue-type plasminogen activator and its expression in Escherichia coli. Mol Biol Med 3(3):279–92PubMedGoogle Scholar
  80. Hart RA, Rinas U, Bailey JE (1990) Protein composition of Vitreoscilla hemoglobin inclusion bodies produced in Escherichia coli. J Biol Chem 265(21):12728–33PubMedGoogle Scholar
  81. Hartl FU, Hayer-Hartl M (2002) Molecular chaperones in the cytosol: from nascent chain to folded protein. Science 295(5561):1852–8PubMedGoogle Scholar
  82. Hartley DL, Kane JF (1988) Properties of inclusion bodies from recombinant Escherichia coli. Biochem Soc Trans 16(2):101–2PubMedGoogle Scholar
  83. Haslbeck M (2002) sHsps and their role in the chaperone network. Cell Mol Life Sci 59(10): 1649–57PubMedGoogle Scholar
  84. Hennessy F, Nicoll WS, Zimmermann R et al. (2005) Not all J domains are created equal: implications for the specificity of Hsp40-Hsp70 interactions. Protein Sci 14(7):1697–709PubMedGoogle Scholar
  85. Hoffmann F, Rinas U (2000) Kinetics of heat-shock response and inclusion body formation during temperature-induced production of basic fibroblast growth factor in high-cell-density cultures of recombinant Escherichia coli. Biotechnol Prog 16(6):1000–7PubMedGoogle Scholar
  86. Hoffmann F, Rinas U (2001) On-line estimation of the metabolic burden resulting from the synthesis of plasmid-encoded and heat-shock proteins by monitoring respiratory energy generation. Biotechnol Bioeng 76(4):333–40PubMedGoogle Scholar
  87. Hoffmann F, Rinas U (2004) Roles of heat-shock chaperones in the production of recombinant proteins in Escherichia coli. Adv Biochem Eng Biotechnol 89:143–61PubMedGoogle Scholar
  88. Hoskins JR, Pak M, Maurizi MR et al. (1998) The role of the ClpA chaperone in proteolysis by ClpAP. Proc Natl Acad Sci USA 95(21):12135–40PubMedGoogle Scholar
  89. Huang GC, Li ZY, Zhou JM et al. (2000) Assisted folding of D-glyceraldehyde-3-phosphate dehydrogenase by trigger factor. Protein Sci 9(6):1254–61PubMedGoogle Scholar
  90. Hunke S, Betton JM (2003) Temperature effect on inclusion body formation and stress response in the periplasm of Escherichia coli. Mol Microbiol 50(5):1579–89PubMedGoogle Scholar
  91. Hunt C, Morimoto RI (1985) Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc Natl Acad Sci USA 82(19): 6455–9PubMedGoogle Scholar
  92. Ivanova MI, Sawaya MR, Gingery M et al. (2004) An amyloid-forming segment of beta2-microglobulin suggests a molecular model for the fibril. Proc Natl Acad Sci USA 101(29):10584–9PubMedGoogle Scholar
  93. Jackson SE (1998) How do small single-domain proteins fold? Fold Des 3(4):R81–91PubMedGoogle Scholar
  94. Jevsevar S, Gaberc-Porekar V, Fonda I et al. (2005) Production of nonclassical inclusion bodies from which correctly folded protein can be extracted. Biotechnol Prog 21(2):632–9PubMedGoogle Scholar
  95. Jungbauer A, Kaar W (2007) Current status of technical protein refolding. J Biotechnol 128(3):587–96PubMedGoogle Scholar
  96. Jurgen B, Lin HY, Riemschneider S et al. (2000) Monitoring of genes that respond to overproduction of an insoluble recombinant protein in Escherichia coli glucose-limited fed-batch fermentations. Biotechnol Bioeng 70(2):217–24PubMedGoogle Scholar
  97. Kapust RB, Tozser J, Copeland TD et al. (2002) The P1 specificity of tobacco etch virus protease. Biochem Biophys Res Commun 294(5):949–55PubMedGoogle Scholar
  98. Kapust RB, Tozser J, Fox JD et al. (2001) Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Protein Eng 14(12): 993–1000PubMedGoogle Scholar
  99. Karplus M (1997) The Levinthal paradox: yesterday and today. Fold Des 2(4):S69–75PubMedGoogle Scholar
  100. Kazemi-Esfarjani P, Benzer S (2000) Genetic suppression of polyglutamine toxicity in Drosophila. Science 287(5459):1837–40PubMedGoogle Scholar
  101. Kedzierska S, Staniszewska M, Wegrzyn A et al. (1999) The role of DnaK/DnaJ and GroEL/GroES systems in the removal of endogenous proteins aggregated by heat-shock from Escherichia coli cells. FEBS Lett 446(2–3):331–7PubMedGoogle Scholar
  102. Kerner MJ, Naylor DJ, Ishihama Y et al. (2005) Proteome-wide analysis of chaperonin-dependent protein folding in Escherichia coli. Cell 122(2):209–20PubMedGoogle Scholar
  103. Khan F, Chuang JI, Gianni S et al. (2003) The kinetic pathway of folding of barnase. J Mol Biol 333(1):169–86PubMedGoogle Scholar
  104. Kitagawa M, Miyakawa M, Matsumura Y et al. (2002) Escherichia coli small heat shock proteins, IbpA and IbpB, protect enzymes from inactivation by heat and oxidants. Eur J Biochem 269(12):2907–17PubMedGoogle Scholar
  105. Krobitsch S, Lindquist S (2000) Aggregation of huntingtin in yeast varies with the length of the polyglutamine expansion and the expression of chaperone proteins. Proc Natl Acad Sci USA 97(4):1589–94PubMedGoogle Scholar
  106. Kuczynska-Wisnik D, Kedzierska S, Matuszewska E et al. (2002) The Escherichia coli small heat-shock proteins IbpA and IbpB prevent the aggregation of endogenous proteins denatured in vivo during extreme heat shock. Microbiology 148(Pt 6):1757–65PubMedGoogle Scholar
  107. Kuczynska-Wisnik D, Zurawa-Janicka D, Narkiewicz J et al. (2004) Escherichia coli small heat shock proteins IbpA/B enhance activity of enzymes sequestered in inclusion bodies. Acta Biochim Pol 51(4):925–31PubMedGoogle Scholar
  108. Laskey RA, Honda BM, Mills AD et al. (1978) Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA. Nature 275(5679):416–20PubMedGoogle Scholar
  109. Laskowska E, Bohdanowicz J, Kuczynska-Wisnik D et al. (2004) Aggregation of heat-shock-denatured, endogenous proteins and distribution of the IbpA/B and Fda marker-proteins in Escherichia coli WT and grpE280 cells. Microbiology 150(Pt 1):247–59PubMedGoogle Scholar
  110. Lemaux PG, Herendeen SL, Bloch PL et al. (1978) Transient rates of synthesis of individual polypeptides in E. coli following temperature shifts. Cell 13(3):427–34PubMedGoogle Scholar
  111. Lethanh H, Neubauer P, Hoffmann F (2005) The small heat-shock proteins IbpA and IbpB reduce the stress load of recombinant Escherichia coli and delay degradation of inclusion bodies. Microb Cell Fact 4(1):6PubMedGoogle Scholar
  112. Levchenko I, Luo L, Baker TA (1995) Disassembly of the Mu transposase tetramer by the ClpX chaperone. Genes Dev 9(19):2399–408PubMedGoogle Scholar
  113. Lewandowska A, Matuszewska M, Liberek K (2007) Conformational properties of aggregated polypeptides determine ClpB-dependence in the disaggregation process. J Mol Biol 371(3):800–11PubMedGoogle Scholar
  114. Lindquist S, Craig EA (1988) The heat-shock proteins. Annu Rev Genet 22:631–77PubMedGoogle Scholar
  115. Malki A, Kern R, Abdallah J et al. (2003) Characterization of the Escherichia coli YedU protein as a molecular chaperone. Biochem Biophys Res Commun 301(2):430–6PubMedGoogle Scholar
  116. Marston FA (1986) The purification of eukaryotic polypeptides synthesized in Escherichia coli. Biochem J 240(1):1–12PubMedGoogle Scholar
  117. Marston FA, Hartley DL (1990) Solubilization of protein aggregates. Methods Enzymol 182: 264–76PubMedGoogle Scholar
  118. Marston FAO, Lowe PA, Doel MT et al. (1984) Purification of Calf Prochymosin(Prorennin) Synthesized in Escherichia coli. Bio/Technology 2(9):800–804Google Scholar
  119. Martinez-Alonso M, Vera A, Villaverde A (2007) Role of the chaperone DnaK in protein solubility and conformational quality in inclusion body-forming Escherichia coli cells. FEMS Microbiol Lett 273(2):187–95PubMedGoogle Scholar
  120. Martinez-Alonso M, Gonz’alez-Montalb’an N, Garcia-Fruit’os E, Villaverde A. (2008) The functional quality of soluble recombinant polypeptides produced in Escherichia coil is defined by a wide conformational spectrum. Appl Environ Microbiol 74(23):7431–3Google Scholar
  121. Matagne A, Dobson CM (1998) The folding process of hen lysozyme: a perspective from the ‘new view’. Cell Mol Life Sci 54(4):363–71PubMedGoogle Scholar
  122. Matouschek A (2003) Protein unfolding–an important process in vivo? Curr Opin Struct Biol 13(1):98–109Google Scholar
  123. Matuszewska M, Kuczynska-Wisnik D, Laskowska E et al. (2005) The small heat shock protein IbpA of Escherichia coli cooperates with IbpB in stabilization of thermally aggregated proteins in a disaggregation competent state. J Biol Chem 280(13):12292–8PubMedGoogle Scholar
  124. Maurizi MR (1992) Proteases and protein degradation in Escherichia coli. Experientia 48(2): 178–201PubMedGoogle Scholar
  125. Middelberg AP (2002) Preparative protein refolding. Trends Biotechnol 20(10):437–43PubMedGoogle Scholar
  126. Miot M, Betton JM (2004) Protein quality control in the bacterial periplasm. Microb Cell Fact 3(1):4PubMedGoogle Scholar
  127. Missiakas D, Schwager F, Betton JM et al. (1996) Identification and characterization of HsIV HsIU (ClpQ ClpY) proteins involved in overall proteolysis of misfolded proteins in Escherichia coli. Embo J 15(24):6899–909PubMedGoogle Scholar
  128. Mogk A, Bukau B (2004) Molecular chaperones: structure of a protein disaggregase. Curr Biol 14(2):R78–80PubMedGoogle Scholar
  129. Mogk A, Deuerling E, Vorderwulbecke S et al. (2003) Small heat shock proteins, ClpB and the DnaK system form a functional triade in reversing protein aggregation. Mol Microbiol 50(2):585–95PubMedGoogle Scholar
  130. Mogk A, Tomoyasu T, Goloubinoff P et al. (1999) Identification of thermolabile Escherichia coli proteins: prevention and reversion of aggregation by DnaK and ClpB. Embo J 18(24): 6934–49PubMedGoogle Scholar
  131. Morell M, Bravo R, Espargaro A et al. (2008): Inclusion bodies: Specificity in their aggregation process and amyloid-like structure. Biochim Biophys Acta 1783(10):1815–25Google Scholar
  132. Morita MT, Kanemori M, Yanagi H et al. (2000) Dynamic interplay between antagonistic pathways controlling the sigma 32 level in Escherichia coli. Proc Natl Acad Sci U S A 97(11):5860–5PubMedGoogle Scholar
  133. Motohashi K, Watanabe Y, Yohda M et al. (1999) Heat-inactivated proteins are rescued by the DnaK.J-GrpE set and ClpB chaperones. Proc Natl Acad Sci USA 96(13):7184–9PubMedGoogle Scholar
  134. Muchowski PJ, Schaffar G, Sittler A et al. (2000) Hsp70 and hsp40 chaperones can inhibit self-assembly of polyglutamine proteins into amyloid-like fibrils. Proc Natl Acad Sci USA 97(14):7841–6PubMedGoogle Scholar
  135. Mujacic M, Bader MW, Baneyx F (2004) Escherichia coli Hsp31 functions as a holding chaperone that cooperates with the DnaK-DnaJ-GrpE system in the management of protein misfolding under severe stress conditions. Mol Microbiol 51(3):849–59PubMedGoogle Scholar
  136. Muramatsu N, Minton AP (1988) Tracer diffusion of globular proteins in concentrated protein solutions. Proc Natl Acad Sci USA 85(9):2984–8PubMedGoogle Scholar
  137. Nagai H, Yuzawa H, Kanemori M et al. (1994) A distinct segment of the sigma 32 polypeptide is involved in DnaK-mediated negative control of the heat shock response in Escherichia coli. Proc Natl Acad Sci USA 91(22):10280–4PubMedGoogle Scholar
  138. Nahalka J (2008) Physiological aggregation of maltodextrin phosphorylase from Pyrococcus furiosus and its application in a process of batch starch degradation to alpha-D-glucose-1-phosphate. J Ind Microbiol Biotechnol 35(4):219–23PubMedGoogle Scholar
  139. Nahalka J, Gemeiner P, Bucko M et al. (2006) Bioenergy beads: a tool for regeneration of ATP/NTP in biocatalytic synthesis. Artif Cells Blood Substit Immobil Biotechnol 34(5):515–21PubMedGoogle Scholar
  140. Nahalka J, Nidetzky B (2007) Fusion to a pull-down domain: a novel approach of producing Trigonopsis variabilisD-amino acid oxidase as insoluble enzyme aggregates. Biotechnol Bioeng 97(3):454–61PubMedGoogle Scholar
  141. Nahalka J, Vikartovska A, Hrabarova E (2008) A crosslinked inclusion body process for sialic acid synthesis. J Biotechnol 134(1–2):146–53Google Scholar
  142. Narberhaus F (2002) Alpha-crystallin-type heat shock proteins: socializing minichaperones in the context of a multichaperone network. Microbiol Mol Biol Rev 66(1):64–93; table of contentsPubMedGoogle Scholar
  143. Neidhardt FC, Umbarger HE (1996) Chemical composition of Escherichia coli. In: (ed) Escherichia coli and Salmonella, American Society for Microbiology Press, Washington, D.C.Google Scholar
  144. Neubauer A, Soini J, Bollok M et al. (2007) Fermentation process for tetrameric human collagen prolyl 4-hydroxylase in Escherichia coli: improvement by gene optimisation of the PDI/beta subunit and repeated addition of the inducer anhydrotetracycline. J Biotechnol 128(2):308–21PubMedGoogle Scholar
  145. Niiranen L, Espelid S, Karlsen CR et al. (2007) Comparative expression study to increase the solubility of cold adapted Vibrio proteins in Escherichia coli. Protein Expr Purif 52(1):210–8PubMedGoogle Scholar
  146. Nishihara K, Kanemori M, Kitagawa M et al. (1998) Chaperone coexpression plasmids: differential and synergistic roles of DnaK-DnaJ-GrpE and GroEL-GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli. Appl Environ Microbiol 64(5):1694–9PubMedGoogle Scholar
  147. Nishihara K, Kanemori M, Yanagi H et al. (2000) Overexpression of trigger factor prevents aggregation of recombinant proteins in Escherichia coli. Appl Environ Microbiol 66(3):884–9PubMedGoogle Scholar
  148. Oberg K, Chrunyk BA, Wetzel R et al. (1994) Nativelike secondary structure in interleukin-1 beta inclusion bodies by attenuated total reflectance FTIR. Biochemistry 33(9):2628–34PubMedGoogle Scholar
  149. Otzen DE, Fersht AR (1998) Folding of circular and permuted chymotrypsin inhibitor 2: retention of the folding nucleus. Biochemistry 37(22):8139–46PubMedGoogle Scholar
  150. Panchenko AR, Luthey-Schulten Z, Wolynes PG (1996) Foldons, protein structural modules, and exons. Proc Natl Acad Sci USA 93(5):2008–13PubMedGoogle Scholar
  151. Parsell DA, Kowal AS, Singer MA et al. (1994) Protein disaggregation mediated by heat-shock protein Hsp104. Nature 372(6505):475–8PubMedGoogle Scholar
  152. Paul DC, Van Frank RM, Muth WL et al. (1983) Immunocytochemical demonstration of human proinsulin chimeric polypeptide within cytoplasmic inclusion bodies of Escherichia coli. Eur J Cell Biol 31(2):171–4PubMedGoogle Scholar
  153. Plakoutsi G, Bemporad F, Calamai M et al. (2005) Evidence for a mechanism of amyloid formation involving molecular reorganisation within native-like precursor aggregates. J Mol Biol 351(4):910–22PubMedGoogle Scholar
  154. Proudfoot AE, Goffin L, Payton MA et al. (1996) In vivo and in vitro folding of a recombinant metalloenzyme, phosphomannose isomerase. Biochem J 318 (Pt 2):437–42Google Scholar
  155. Przybycien TM, Dunn JP, Valax P et al. (1994) Secondary structure characterization of beta-lactamase inclusion bodies. Protein Eng 7(1):131–6PubMedGoogle Scholar
  156. Rajan RS, Illing ME, Bence NF et al. (2001) Specificity in intracellular protein aggregation and inclusion body formation. Proc Natl Acad Sci USA 98(23):13060–5PubMedGoogle Scholar
  157. Ranson NA, Farr GW, Roseman AM et al. (2001) ATP-bound states of GroEL captured by cryo-electron microscopy. Cell 107(7):869–79PubMedGoogle Scholar
  158. Rinas U, Bailey JE (1992) Protein compositional analysis of inclusion bodies produced in recombinant Escherichia coli. Appl Microbiol Biotechnol 37(5):609–14PubMedGoogle Scholar
  159. Rinas U, Bailey JE (1993) Overexpression of bacterial hemoglobin causes incorporation of pre-beta-lactamase into cytoplasmic inclusion bodies. Appl Environ Microbiol 59(2):561–6PubMedGoogle Scholar
  160. Rinas U, Boone TC, Bailey JE (1993) Characterization of inclusion bodies in recombinant Escherichia coli producing high levels of porcine somatotropin. J Biotechnol 28(2–3):313–20PubMedGoogle Scholar
  161. Rinas U, Hoffmann F, Betiku E et al. (2007) Inclusion body anatomy and functioning of chaperone-mediated in vivo inclusion body disassembly during high-level recombinant protein production in Escherichia coli. J Biotechnol 127(2):244–57PubMedGoogle Scholar
  162. Rosen R, Biran D, Gur E et al. (2002) Protein aggregation in Escherichia coli: role of proteases. FEMS Microbiol Lett 207(1):9–12PubMedGoogle Scholar
  163. Rudolph R, Lilie H (1996) In vitro folding of inclusion body proteins. Faseb J 10(1):49–56PubMedGoogle Scholar
  164. Sahdev S, Khattar SK, Saini KS (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies. Mol Cell Biochem 307(1–2):249–64PubMedGoogle Scholar
  165. Sakikawa C, Taguchi H, Makino Y et al. (1999) On the maximum size of proteins to stay and fold in the cavity of GroEL underneath GroES. J Biol Chem 274(30):21251–6PubMedGoogle Scholar
  166. Sastry MS, Korotkov K, Brodsky Y et al. (2002) Hsp31, the Escherichia coli yedU gene product, is a molecular chaperone whose activity is inhibited by ATP at high temperatures. J Biol Chem 277(48):46026–34PubMedGoogle Scholar
  167. Schein CH, Noteborn (1988) Formation of soluble recombinant proteins in Escherichia coli is favored by lower growth temperature. Bio/Technology 6(3):291–294Google Scholar
  168. Schirmer EC, Glover JR, Singer MA et al. (1996) HSP100/Clp proteins: a common mechanism explains diverse functions. Trends Biochem Sci 21(8):289–96PubMedGoogle Scholar
  169. Schlieker C, Tews I, Bukau B et al. (2004) Solubilization of aggregated proteins by ClpB/DnaK relies on the continuous extraction of unfolded polypeptides. FEBS Lett 578(3):351–6PubMedGoogle Scholar
  170. Schmidt M, Babu KR, Khanna N et al. (1999) Temperature-induced production of recombinant human insulin in high-cell density cultures of recombinant Escherichia coli. J Biotechnol 68(1):71–83PubMedGoogle Scholar
  171. Schoemaker JM, Brasnett AH, Marston FA (1985) Examination of calf prochymosin accumulation in Escherichia coli: disulphide linkages are a structural component of prochymosin-containing inclusion bodies. Embo J 4(3):775–80PubMedGoogle Scholar
  172. Schoner RG, Ellis LF, Schoner BE (1985) Isolation and purification of protein granules from Escherichia coli cells overproducing bovine growth hormone. Bio/Technology 3:151–154Google Scholar
  173. Schrodel A, de Marco A (2005) Characterization of the aggregates formed during recombinant protein expression in bacteria. BMC Biochem 6:10PubMedGoogle Scholar
  174. Schultz T, Martinez L, de Marco A (2006) The evaluation of the factors that cause aggregation during recombinant expression in E. coli is simplified by the employment of an aggregation-sensitive reporter. Microb Cell Fact 5:28PubMedGoogle Scholar
  175. Shearstone JR, Baneyx F (1999) Biochemical characterization of the small heat shock protein IbpB from Escherichia coli. J Biol Chem 274(15):9937–45PubMedGoogle Scholar
  176. Snow CD, Nguyen H, Pande VS et al. (2002) Absolute comparison of simulated and experimental protein-folding dynamics. Nature 420(6911):102–6PubMedGoogle Scholar
  177. Sorensen HP, Mortensen KK (2005a) Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol 115(2):113–28PubMedGoogle Scholar
  178. Sorensen HP, Mortensen KK (2005b) Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli. Microb Cell Fact 4(1):1PubMedGoogle Scholar
  179. Speed MA, Wang DI, King J (1996) Specific aggregation of partially folded polypeptide chains: the molecular basis of inclusion body composition. Nat Biotechnol 14(10):1283–7PubMedGoogle Scholar
  180. Spiess C, Beil A, Ehrmann M (1999) A temperature-dependent switch from chaperone to protease in a widely conserved heat shock protein. Cell 97(3):339–47PubMedGoogle Scholar
  181. Stefani M, Dobson CM (2003) Protein aggregation and aggregate toxicity: new insights into protein folding, misfolding diseases and biological evolution. J Mol Med 81(11):678–99PubMedGoogle Scholar
  182. Straus DB, Walter WA, Gross CA (1987) The heat shock response of E. coli is regulated by changes in the concentration of sigma 32. Nature 329(6137):348–51PubMedGoogle Scholar
  183. Sugrue R, Marston FA, Lowe PA et al. (1990) Denaturation studies on natural and recombinant bovine prochymosin (prorennin). Biochem J 271(2):541–7PubMedGoogle Scholar
  184. Surewicz WK, Mantsch HH (1988) New insight into protein secondary structure from resolution-enhanced infrared spectra. Biochim Biophys Acta 952(2):115–30PubMedGoogle Scholar
  185. Surewicz WK, Stepanik TM, Szabo AG et al. (1988) Lipid-induced changes in the secondary structure of snake venom cardiotoxins. J Biol Chem 263(2):786–90PubMedGoogle Scholar
  186. Teter SA, Houry WA, Ang D et al. (1999) Polypeptide flux through bacterial Hsp70: DnaK cooperates with trigger factor in chaperoning nascent chains. Cell 97(6):755–65PubMedGoogle Scholar
  187. Thomas JG, Ayling A, Baneyx F (1997) Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins from E. coli. To fold or to refold. Appl Biochem Biotechnol 66(3):197–238PubMedGoogle Scholar
  188. Thomas JG, Baneyx F (1996a) Protein folding in the cytoplasm of Escherichia coli: requirements for the DnaK-DnaJ-GrpE and GroEL-GroES molecular chaperone machines. Mol Microbiol 21(6):1185–96PubMedGoogle Scholar
  189. Thomas JG, Baneyx F (1996b) Protein misfolding and inclusion body formation in recombinant Escherichia coli cells overexpressing Heat-shock proteins. J Biol Chem 271(19):11141–7PubMedGoogle Scholar
  190. Thomas JG, Baneyx F (1998) Roles of the Escherichia coli small heat shock proteins IbpA and IbpB in thermal stress management: comparison with ClpA, ClpB, and HtpG In vivo. J Bacteriol 180(19):5165–72PubMedGoogle Scholar
  191. Thomas JG, Baneyx F (2000) ClpB and HtpG facilitate de novo protein folding in stressed Escherichia coli cells. Mol Microbiol 36(6):1360–70PubMedGoogle Scholar
  192. Tokatlidis K, Dhurjati P, Millet J et al. (1991) High activity of inclusion bodies formed in Escherichia coli overproducing Clostridium thermocellum endoglucanase D. FEBS Lett 282(1):205–8PubMedGoogle Scholar
  193. Tomoyasu T, Arsene F, Ogura T et al. (2001a) The C terminus of sigma(32) is not essential for degradation by FtsH. J Bacteriol 183(20):5911–7PubMedGoogle Scholar
  194. Tomoyasu T, Mogk A, Langen H et al. (2001b) Genetic dissection of the roles of chaperones and proteases in protein folding and degradation in the Escherichia coli cytosol. Mol Microbiol 40(2):397–413PubMedGoogle Scholar
  195. Tomoyasu T, Ogura T, Tatsuta T et al. (1998) Levels of DnaK and DnaJ provide tight control of heat shock gene expression and protein repair in Escherichia coli. Mol Microbiol 30(3):567–81PubMedGoogle Scholar
  196. Tsumoto K, Umetsu M, Kumagai I et al. (2003) Solubilization of active green fluorescent protein from insoluble particles by guanidine and arginine. Biochem Biophys Res Commun 312(4):1383–6PubMedGoogle Scholar
  197. Umetsu M, Tsumoto K, Ashish K et al. (2004) Structural characteristics and refolding of in vivo aggregated hyperthermophilic archaeon proteins. FEBS Lett 557(1–3):49–56PubMedGoogle Scholar
  198. Umetsu M, Tsumoto K, Nitta S et al. (2005) Nondenaturing solubilization of beta2 microglobulin from inclusion bodies by L-arginine. Biochem Biophys Res Commun 328(1):189–97PubMedGoogle Scholar
  199. Vallejo LF, Rinas U (2004) Strategies for the recovery of active proteins through refolding of bacterial inclusion body proteins. Microb Cell Fact 3(1):11PubMedGoogle Scholar
  200. Veinger L, Diamant S, Buchner J et al. (1998) The small heat-shock protein IbpB from Escherichia coli stabilizes stress-denatured proteins for subsequent refolding by a multichaperone network. J Biol Chem 273(18):11032–7PubMedGoogle Scholar
  201. Vendruscolo M, Paci E, Dobson CM et al. (2003) Rare fluctuations of native proteins sampled by equilibrium hydrogen exchange. J Am Chem Soc 125(51):15686–7PubMedGoogle Scholar
  202. Ventura S (2005) Sequence determinants of protein aggregation: tools to increase protein solubility. Microb Cell Fact 4(1):11PubMedGoogle Scholar
  203. Ventura S, Villaverde A (2006) Protein quality in bacterial inclusion bodies. Trends Biotechnol 24(4):179–85PubMedGoogle Scholar
  204. Ventura S, Zurdo J, Narayanan S et al. (2004) Short amino acid stretches can mediate amyloid formation in globular proteins: the Src homology 3 (SH3) case. Proc Natl Acad Sci USA 101(19):7258–63PubMedGoogle Scholar
  205. Vera A, Aris A, Carrio M et al. (2005) Lon and ClpP proteases participate in the physiological disintegration of bacterial inclusion bodies. J Biotechnol 119(2):163–71PubMedGoogle Scholar
  206. Vera A, Gonzalez-Montalban N, Aris A et al. (2007) The conformational quality of insoluble recombinant proteins is enhanced at low growth temperatures. Biotechnol Bioeng 96(6):1101–6PubMedGoogle Scholar
  207. Villaverde A, Carrio MM (2003) Protein aggregation in recombinant bacteria: biological role of inclusion bodies. Biotechnol Lett 25(17):1385–95PubMedGoogle Scholar
  208. Wagner S, Baars L, Ytterberg AJ et al. (2007) Consequences of membrane protein overexpression in Escherichia coli. Mol Cell Proteomics 6(9):1527–50PubMedGoogle Scholar
  209. Waldo GS, Standish BM, Berendzen J et al. (1999) Rapid protein-folding assay using green fluorescent protein. Nat Biotechnol 17(7):691–5PubMedGoogle Scholar
  210. Walsh G (2005) Therapeutic insulins and their large-scale manufacture. Appl Microbiol Biotechnol 67(2):151–9PubMedGoogle Scholar
  211. Wang L, Maji SK, Sawaya MR et al. (2008): Bacterial inclusion bodies contain amyloid-like strucure. PLoS Biol 6(8):e195Google Scholar
  212. Chan HY, Gray-Board GL et al. (1999) Suppression of polyglutamine-mediated neurodegeneration in Drosophila by the molecular chaperone HSP70. Nat Genet 23(4):425–8PubMedGoogle Scholar
  213. Wickner S, Gottesman S, Skowyra D et al. (1994) A molecular chaperone, ClpA, functions like DnaK and DnaJ. Proc Natl Acad Sci USA 91(25):12218–22PubMedGoogle Scholar
  214. Wickner S, Maurizi MR, Gottesman S (1999) Posttranslational quality control: folding, refolding, and degrading proteins. Science 286(5446):1888–93PubMedGoogle Scholar
  215. Williams DC, Van Frank RM, Muth WL et al. (1982) Cytoplasmic inclusion bodies in Escherichia coli producing biosynthetic human insulin proteins. Science 215(4533):687–9PubMedGoogle Scholar
  216. Wolynes PG, Onuchic JN, Thirumalai D (1995) Navigating the folding routes. Science 267(5204):1619–20PubMedGoogle Scholar
  217. Worrall DM, Goss NH (1989) The formation of biologically active beta-galactosidase inclusion bodies in Escherichia coli. Aust J Biotechnol 3(1):28–32PubMedGoogle Scholar
  218. Yokoyama K, Kikuchi Y, Yasueda H (1998) Overproduction of DnaJ in Escherichia coli improves in vivo solubility of the recombinant fish-derived transglutaminase. Biosci Biotechnol Biochem 62(6):1205–10PubMedGoogle Scholar
  219. Yon JM (2001) Protein folding: a perspective for biology, medicine and biotechnology. Braz J Med Biol Res 34(4):419–35PubMedGoogle Scholar
  220. Young JC, Agashe VR, Siegers K et al. (2004) Pathways of chaperone-mediated protein folding in the cytosol. Nat Rev Mol Cell Biol 5(10):781–91PubMedGoogle Scholar
  221. Yura T, Nakahigashi K (1999) Regulation of the heat-shock response. Curr Opin Microbiol 2(2):153–8PubMedGoogle Scholar
  222. Zhang YB, Howitt J, McCorkle S et al. (2004) Protein aggregation during overexpression limited by peptide extensions with large net negative charge. Protein Expr Purif 36(2):207–16PubMedGoogle Scholar
  223. Zhu X, Zhao X, Burkholder WF et al. (1996) Structural analysis of substrate binding by the molecular chaperone DnaK. Science 272(5268):1606–14PubMedGoogle Scholar
  224. Zimmerman SB, Minton AP (1993) Macromolecular crowding: biochemical, biophysical, and physiological consequences. Annu Rev Biophys Biomol Struct 22:27–65PubMedGoogle Scholar
  225. Zimmerman SB, Trach SO (1991) Estimation of macromolecule concentrations and excluded volume effects for the cytoplasm of Escherichia coli. J Mol Biol 222(3):599–620PubMedGoogle Scholar
  226. Zolkiewski M (1999) ClpB cooperates with DnaK, DnaJ, and GrpE in suppressing protein aggregation. A novel multi-chaperone system from Escherichia coli. J Biol Chem 274(40):28083–6PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Elena Garcìa-Fruitòs
  • Nuria Gonzàlez-Montalbàn
  • Mònica Martìnez-Alonso
  • Ursula Rinas
  • Antonio Villaverde
    • 1
  1. 1.Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de BarcelonaBellaterra08193 Spain

Personalised recommendations