Part:BBa_K538001

Cpn60 (O. antarctica)

Figure 1: (a) E. coli 's growth rate as a function of temperature, in strains that do (white circles) or do not (black circles) express Cpn60/10, and (b) in vitro refolding activities of Cpn60/10 as a function of temperature (black circles), compared with that of E. coli 's endogenous GroEL/ES (white circles). Adapted from Ferrer et al. (2003)

This BioBrick encodes the protein chaperonin 60, which is part of the Cpn60/10 chaperone system of Oleispira antarctica. When coexpressed with cochaperonin 10 (Cpn10, BBa_K538000), it forms a heteromultimer that can refold enzymes at very low temperatures, thus safeguarding their functionality. This has been shown to enable E. coli to grow even at freezing point. (Figure 1)

Usage and Biology

Cpn10 and Cpn60 are, respectively, homologous to GroES and GroEL of E. coli. The GroEL/ES chaperone system promotes the folding and/or assembly of over 30% of E. coli's cellular proteins, is required for bacteriophage morphogenesis and has a role in protein secretion.[http://www.nature.com/nature/journal/v355/n6355/abs/355033a0.html][http://onlinelibrary.wiley.com/doi/10.1002/1521-3773%2820020402%2941:7%3C1098::AID-ANIE1098%3E3.0.CO;2-9/abstract] However, it rapidly loses its refolding activity at temperatures below 37°C. (Figure 1b)[http://www.nature.com/nbt/journal/v21/n11/full/nbt1103-1266b.html] Cpn60/10, on the other hand, functions very well at these temperatures.

Ferrer et al. demonstrated that Cpn60's product, chaperonin 60, coprecipitates with many of the genes found in the proteome of E. coli.[http://onlinelibrary.wiley.com/doi/10.1002/pmic.200500031/abstract] Also, by transforming E. coli with Cpn10 and Cpn60, they've enabled it to grow even at freezing point. (Figure 1a)

[Under construction]

References

1. Gething & Sambrook Protein folding in the cell, Nature 355, 33–45 (1992)
2. Walter & Buchner Molecular Chaperones — Cellular Machines for Protein Folding, Angew. Chem. Int. Ed. Eng. 41, 1098–1113 (2002)
3. Ferrer et al. Chaperonins govern growth of Escherichia coli at low temperatures, Nat. Biotech. 21, 1266 - 1267 (2003)
4. Strocchi, Ferrer, Timmis & Golyshin Low temperature-induced systems failure in Escherichia coli: Insights from rescue by cold-adapted chaperones, Proteomics 6 (1), 193-206 (2005)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 870
  Illegal BamHI site found at 936
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]