Fig. 4.

Sis1 (CTDI)-Ssa1 (EEVD) interaction is crucial for the superior, aggregate-remodeling activity of Sis1-Ssa1. (A) Binding of chaperone variants (as color indicated in the legend) to luciferase aggregates immobilized on the BLI sensor, according to the scheme in Fig. 1B. (B) Binding of Sis1 E50A and Ssa1 ΔEEVD to luciferase aggregates cross-linked with glutaraldehyde (orange) and nonmodified (yellow), as on the scheme in Fig. 2C. The presented results are representative for three replicates. (C) Luciferase reactivation by the system upon sequential addition of different chaperone variants (as color indicated in the legend), according to the scheme in Fig. 3C. Luciferase activity was normalized to the native protein. Error bars show SD from three independent repeats. Dashed lines indicate the starting points of the subsequent steps of experiments.