Fig. 5.

Mycolactone efficiently blocks type II TMP integration. (A) Full-length Ii (wild type and G47L Q48L mutant) and the Ii₁₂₅ truncation used in this study. (B) Estimated TMD hydrophobicities (kcal/mol) of Ii and Ii_{G47L Q48L}. (C) Graph shows the reduction in the amount of glycosylated (+g) Ii and related constructs in the presence of mycolactone (MYC), relative to control samples as described in the legend to Fig. 1. The statistical test performed was one-way ANOVA. Error bars show mean±s.d. (n=3). P-values are as defined in Fig. 1 legend. Translation in the absence or presence of mycolactone performed using Ii (D), Ii_{G47L Q48L} (E) and Ii₁₂₅ (F), which was followed by treatment with EndoglycosidaseH (EndoH). (G) Ii truncations used in this study. For crosslinking experiments, truncations contained either a native cysteine residue (C28) or one that was artificially introduced [*(50)]. A truncated version of TNFα used for crosslinking analysis (as described in MacKinnon et al., 2014) is shown for comparative purposes. Crosslinking was performed on Ii truncations (H) and Ii₁₂₅*(50) (I) and the resulting adducts are labelled as described in the Fig. 4G legend. Other symbols are as defined in Fig. 1 legend. Puro, puromycin.