Comparison of the conformational state and in vitro refolding of yeast chaperonin protein cpn10 with bacterial GroES.

Gel filtration studies demonstrate that the heptameric complex of yeast cpn10 (pI around 8.8) reversibly disassembles into monomers when lowering the pH to 4.5, whereas its secondary structure is retained as demonstrated by circular dichroism. Monomeric yeast cpn10 does not bind to GroEL in the presence of nucleotides, whereas under identical conditions E. coli cpn10 (GroES), having a strong sequence homology to the yeast form but a pI of 5.2, shows no pH-dependent dissociation and is able to complex with GroEL at both pH 7.5 and 4.5. Using circular dichroism it is shown that, unlike E. coli cpn10, yeast cpn10 is not able to refold spontaneously after first being fully unfolded in 8 M urea. However, refolding of yeast cpn10 to a complex that can be recognised by GroEL depends on the presence of a lipid-water interface with a specificity for negatively charged lipids. We suggest that the requirements for refolding of yeast cpn10 are related to its post-translational transport and subcellular localization.