Effect of chaperones on reactivation of heat-inactivated LDH. Indicated components were added to the solutions containing LDH (final concentration, 0.2 μM), and the solutions were incubated at 73°C for 30 min. Then the temperature was shifted down to 55°C, and other indicated components were immediately supplemented to the solutions (0 min in abscissa). Incubation at 55°C continued, and LDH activities were measured at the indicated times. Final concentrations of added components were 0.4 μM (TDnaK⋅J complex), 0.8 μM (TGrpE), 0.4 μM (TClpB), 0.4 μM (TGroEL/ES), and 3 mM (nucleotides). Note that components added at 73°C remained present after the temperature shift. The KJ, E, and B represent the TDnaK⋅J complex, TGrpE, and TClpB, respectively. The orders of addition are shown. (A) Effect of the TDnaK⋅J complex and TGrpE. (B) Effect of TClpB. (C) Effect of TDnaK⋅J complex, TGrpE, TClpB, and TGroEL/ES. (D) Effect of nucleotides.