Extended Data Fig. 3: C-terminal truncation abolishes DNAJB1 interaction and disaggregation activity.
From: Molecular dissection of amyloid disaggregation by human HSP70
a, b, Cartoon representations of an α-syn amyloid protofilament (PDB 2N0A42) illustrating the effects of chaperone binding (a, DNAJB1; b, HSP70) on site specific BADAN fluorescence (Fig. 1c, d) introduced at the indicated sites. The colour gradient reflects the strength of the interaction. DNAJB1 binding primarily affects fluorophores introduced at the C terminus, whereas the effects of HSP70 binding are specific to the N terminus. c, Schematic representation of N- and C-terminal truncation constructs of α-syn. d, Immunoblot of insoluble (pellet, P) and soluble (S) fractions of fibrillar α-syn truncation mutants in the absence or presence of chaperones (DNAJB1, HSP70, HSP110) and ATP after 16-h incubation. e, Steady-state fluorescence anisotropy of DNAJB1AF488 after titration of indicated fibrillar C-terminal truncations of α-syn. Binding curves were fitted to a one-site binding model using GraphPad Prism. f, Disaggregation of fibrillar C-terminal truncations of α-syn by the HSP70 chaperone machinery (DNAJB1, HSP70, HSP110) and ATP monitored by ThT fluorescence. Reaction conditions and number of independent replicates are specified in Extended Data Table 2. Data are mean ± s.e.m.
