Fig. 4: The N-terminus of the PtsG-dependent entry domain controls a conformational switch.
From: Proteolytic processing induces a conformational switch required for antibacterial toxin delivery
a CD spectra of PtsG-dependent entry domains. b 1H-15N HSQC NMR spectra of PtsG-dependent entry domains. c Chemical denaturation of PtsG-dependent entry domains. Purified proteins were denatured in urea and unfolding monitored by CD at 224 nm. d CD spectra of ∆VENN and ∆VENN-Tyr5Ala entry domains. e Chemical denaturation of ∆VENN and ∆VENN-Tyr5Ala entry domains. f Outer-membrane bypass. Purified CdiA-CTEC3006 variants were incubated with polymyxin B (PMB) treated E. coli cells, and total RNA was isolated for Northern blot analysis using a probe to tRNAIle. Where indicated, cells carried the ∆ptsG allele and/or a plasmid that expresses cdiIEC3006. g In vitro nuclease activity. E. coli total RNA was treated with CdiA-CTEC3006 proteins in the absence and presence of purified CdiIEC3006 immunity protein (CdiI-His6). The experiments in panels a, c, e, and g were repeated independently twice with similar results. The experiments in panels b, d, and f were performed once. Source data are provided as a Source Data file.
