Small Heat Shock Proteins Potentiate Amyloid Dissolution by Protein Disaggregases from Yeast and Humans (original) (raw)
Figure 9
Hsp42 synergizes with Ssa1:Sis1 and Ssa1:Ydj1 to inhibit seeded assembly of NM.
(A–E) NM fibers (5 µM NM monomer) were incubated for 60 min at 25°C with buffer or Hsp42 (10 µM). NM fibers were then recovered by centrifugation at 16,000 g, gently washed, resuspended in an equal volume of assembly buffer, and used to seed (2% wt/wt) assembly of NM (5 µM) for 12 h at 25°C in the presence of ATP (5 mM) and increasing concentrations of Ssa1 (A), Sis1 (B), Ydj1 (C), Ssa1:Sis1 (D), or Ssa1:Ydj1 (E). Fibrillization was monitored by ThT fluorescence. Values represent means±SD (n = 3). (F) NM fibers (5 µM or 1.25 µM) were sonicated and transformed into [_psi_−] cells overexpressing the indicated combination of Hsp26, Hsp42, Sis1, and Ydj1. The proportion of [PSI+] colonies was then determined. Values represent means±SD (n = 5).