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. 2020 Jul 2;142(28):12157–12166. doi: 10.1021/jacs.0c02362

Figure 3.

Figure 3

Single-molecule observation of ColE9-IUTD translocation through OmpF. (a) The ColE9-IUTD sequence (residues 2–83) was produced by TEV protease, which cleaves the TEV site (ENLYFQ/GA) between the IUTD and the fusion tag. The locations of OBS1, TBE, and OBS2 are shown in red, purple, and orange with residue numbers. The residual amino acids from the TEV site (ENLYFQ) are shown as a dashed line. (b) Current trace of OmpF when 40 nM free IUTD (ColE92–83-ENLYFQ) was added to the extracellular side of OmpF at −100 mV. The IUTD first reversibly associates with a first OmpF subunit (red asterisk). The latency before the C1 state (τ01) indicates the time before translocation into the first OmpF subunit. The latency before the initial passage of OBS1 into the second subunit is τ12, and the latency before rebinding is τ12′.