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

__NOTOC__
===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/full/nbt1103-1266b.html] (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 ''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) Their findings suggest inactivation of a few cold-sensitive key 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. It is currently being investigated whether or not the CryoBricks comprising [[Part:BBa_K538000 | BBa_K538000]] and [[Part:BBa_K538001 | BBa_K538001]] reproduce this effect.


'''[Update pending; waiting for lab results]'''


In case the chaperones' effect on growth rate at low temperatures '''is''' reproducible, team [http://2011.igem.org/Team:Amsterdam Amsterdam] expects a similar result when transforming related species with their CryoBricks. If systems failure is caused by inactivation of genes like ''Dps'', ''ClpB'', ''DnaK'' and ''RpsB'' (or their homologs), ''Cpn60/10'' might be able to recover their activity. This brick's developers had intended to investigate this in some bacterial species, but constraints on time made this unfeasible.


===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)