Figure 2. Ssa1 efficiently prevents TA aggregation, whereas the currently known GET components do not.

• A, B
  Time courses of Sbh1 (A) and Bos1 (B) aggregation measured by the turbidity assay.
• C, D
  Sedimentation analysis of recombinant Sbh1 (C) and Bos1 (D). 1.5 μM Sbh1 or Bos1 was incubated in assay buffer with or without the indicated factors and ultracentrifuged at 390,880 g for 30 min. Total input (T), soluble (S), and pellet (P) fractions were resolved by SDS–PAGE and visualized by silver stain. 1.5 μM Sgt2 was used in (C), and 3 μM Ssa1 and/or Sgt2 was used in (D) where indicated. The lower panel in (D) shows quantification of soluble Bos1.
• E, F
  Sedimentation analysis of ³⁵S‐labeled Bos1 synthesized by PURE‐IVT. Where indicated, translations reactions were supplemented with 0.5 μM Sgt2, 0.25 μM Get4/5, 0.75 μM Get3, and/or 3 μM Ssa1. The lower panels show quantification of soluble Bos1 under the indicated reaction conditions.
• G
  Get4/5 exacerbates Bos1 aggregation in the turbidity assay. Reactions used 1.5 μM recombinant Bos1, 1.5 μM Sgt2 without (blue) and with (green) 1.5 μM Get4/5 present.
• H
  Time courses of Bos1 aggregation in the presence of Ssa1.
• I
  Ssa1 solubilizes Bos1 in a dose‐dependent manner. The line was a fit of the data to equation (1).

Data information: All values are reported as mean ± SD, with n = 3–4.Source data are available online for this figure.