The guanidine-induced conformational changes of the chaperonin GroEL from Escherichia coli. Evidence for the existence of an unfolding intermediate state.

Equilibrium unfolding experiments of the E. coli chaperonin GroEL were performed in guanidine hydrochloride. A reversible unfolding intermediate was observed in very low concentrations of denaturant (< 0.5 M guanidine hydrochloride). This intermediate was characterized by a decreased light scattering intensity and an increased binding of the fluorescent probe 1-anilino-8-naphthalene sulfonate. No significant changes in circular dichroism spectra were observed for this unfolding intermediate. A second decrease in fluorescence intensity and light scattering was observed in higher concentrations of guanidine hydrochloride, with a transitional midpoint of 1.15 M. This transition was accompanied by the complete loss of secondary structure, as monitored by circular dichroism spectroscopy. This second transition agreed well with the results previously reported in this journal (Price et al. (1993) Biochim. Biophys. Acta 1161, 52-58).