The chaperonin GroEL binds to non-native substrate proteins via hydrophobic interactions, preventing their aggregation, which is minimized at low temperatures. In the present study, we investigated the refolding of urea-denatured rhodanese at low temperatures, in the presence of ox-GroEL (oxidized GroEL), which contains increased exposed hydrophobic surfaces and retains its ability to hydrolyse ATP. We found that ox-GroEL could efficiently bind the urea-unfolded rhodanese at 4°C, without requiring excess amount of chaperonin relative to normal GroEL (i.e. non-oxidized). The release/reactivation of rhodanese from GroEL was minimal at 4°C, but was found to be optimal between 22 and 37°C. It was found that the loss of the ATPase activity of ox-GroEL at 4°C prevented the release of rhodanese from the GroEL–rhodanese complex. Thus ox-GroEL has the potential to efficiently trap recombinant or non-native proteins at 4°C and release them at higher temperatures under appropriate conditions.
Interaction of oxidized chaperonin GroEL with an unfolded protein at low temperatures
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Girish C. Melkani, Robin Sielaff, Gustavo Zardeneta, Jose A. Mendoza; Interaction of oxidized chaperonin GroEL with an unfolded protein at low temperatures. Biosci Rep 1 June 2012; 32 (3): 299–303. doi: https://doi.org/10.1042/BSR20110104
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