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. 2017 Feb 20;36(6):783–796. doi: 10.15252/embj.201593378

Figure EV4. KJE dissociates IbpAB from IbpAB–MDH assemblies in vitro .

Figure EV4

  1. Experimental scheme. IbpAB–MDH assemblies, isolated from unbound IbpAB via sedimentation in glycerol gradient, were incubated with the indicated components for 120 min and resubjected to a second round of sedimentation.
  2. KJE dissociates IbpAB from IbpAB–MDH assemblies. Fractions were collected from the top of the gradient, and MDH and IbpA were visualized by Western blot following SDS–PAGE. IbpA and MDH levels in each fraction were determined by quantitative immunoblot analysis using IbpA‐ and MDH‐specific antibodies, respectively. The band intensities of IbpA and MDH retained in the core assembly (fractions 5–12) or released from the assembly (fractions 1–3) were quantified and plotted on the graphs on the left. IbpAB–MDH assemblies (approximate stoichiometry 1:0.5:1), DnaK, DnaJ, GrpE and ClpB were present at 400 nM (calculated for MDH monomers), 1 μM, 0.3 μM, 0.3 μM and 1.5 μM concentrations, respectively.

Source data are available online for this figure.