Fig. 3: TAM can promote the assembly of OMPs in vitro.

Urea-denatured OmpA (a), EspPΔ5’ (b), Ag43-β (c), or FadL (d) were incubated with PLE proteoliposomes containing 2 μM TAM, TamA, or BAM or empty PLE liposomes at 30 °C. Samples were collected from each reaction after 0.5 and 60 min, treated with proteinase K (PK), or left untreated. After adding the loading buffer, samples were placed on ice or heated to 95 °C and resolved by SDS-PAGE. OMP folding was assessed by determining the fraction of the protein that was resistant to SDS denaturation in the absence of heat (and that underwent autocatalytic cleavage in the case of EspPΔ5’) by Western blot using the indicated antiserum. For clarity, an experiment that shows a more highly resolved view of the folded forms of Ag43-β is depicted in Supplementary Fig. 4b, and an experiment in which the assembly of Ag43-β in the presence of TAM/PLE or TamA/PLE was repeated but the blot was overexposed is shown in Supplementary Fig. 4c. All of the experiments shown here were repeated three times with similar results.