View source for Template:2011/Team:Amsterdam/Cpn60 10

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===Biology===
''Cpn10'' and ''Cpn60'' are homologous to ''GroES'' and ''GroEL'' of ''E. coli'', respectively. 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.[http://www.nature.com/nbt/journal/v21/n11/pdf/nbt1103-1266b.pdf] (Figure 1b) ''Cpn60/10'', on the other hand, functions very well at these temperatures.

Ferrer ''et al.'' identified 22 housekeeping proteins involved with ''E. coli'' 's systems failure at low temperatures. They went on to demonstrate their interactions with chaperones are key determinants of activity at these temperatures. It was shown that the ''Cpn60'' protein from ''O. antarctica'' coprecipitates with many of the proteins 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) Their findings suggest inactivation of a few cold-sensitive key proteins causes systems failure in ''E. coli'', and that cells may be 'rescued' by reactivating these genes.


===Characterisation===
Ferrer ''et al.'' have already shown the ''Cpn60/10'' system to have a nameworthy impact on ''E. coli'''s cold tolerance. Unfortunately, this brick's developers only managed to assemble a protein generator comprising ''Cpn10'' 's coding region. Reproducing the results of Ferrer ''et al.'' would have required coexpression of ''Cpn10'' and ''Cpn60'', so this impossible.

Attempts '''were''' made to characterise whether or not ''Cpn10'' expression can bestow a certain degree of cold resistance independent of ''Cpn60''. According to expectation, ''Cpn10'' doesn't significantly affect ''E. coli'' 's specific growth rate at suboptimal temperatures. Refer to the part's [[Part:BBa_K538000:Experience|experience]] page for the graph summarizing the ''Cpn10'' generator's growth rate investigation.

In addition to investigating the effect of ''Cpn10'' on growth rate a suboptimal temperatures, the effect its expression has on freeze/thaw cycle survival was characterised. During this experiment, a most remarkable observation was made; the data suggests ''Cpn10'' enhances ''E. coli'' 's  freeze/thaw survival rate in and of itself, without being coexpressed with ''Cpn60''. Experiments to verify or falsify this suggestion are currently being prepared and executed. This section will be updated in due time.


===Safety===
There are risks involved with enhancing bacterial cold resistance and facilitating their growth at low temperatures. Please refer to the [http://2011.igem.org/Team:Amsterdam/Project/Safety safety page] of this brick's developers.


===References===
# '''Gething & Sambrook''' Protein folding in the cell, ''Nature 355, 33–45'' (1992)
# '''Walter & Buchner''' Molecular Chaperones — Cellular Machines for Protein Folding, ''Angew. Chem. Int. Ed. Eng. 41, 1098–1113'' (2002)
# '''Ferrer ''et al.''''' Chaperonins govern growth of ''Escherichia coli'' at low temperatures, ''Nat. Biotech. 21, 1266 - 1267'' (2003)
# '''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)