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. 2017 Feb 21;114(10):E1958–E1967. doi: 10.1073/pnas.1615056114

Fig. 4.

Fig. 4.

The TatB suppressors do not restore signal peptide binding to the TatBC complex. (A) C-terminally His-tagged GFP with the wild-type (RR) or twin-arginine substituted SufI signal peptide at its N terminus, as indicated, was purified by magnetic nickel beads from digitonin-treated cell extracts coexpressing TatC along with either wild-type TatB or the E8K, F13Y, or I36N substituted variants. (B) C-terminally His-tagged GFP with the wild-type SufI signal peptide at its N terminus was purified by magnetic nickel beads from digitonin-treated cell extracts coexpressing TatB and TatC with the indicated amino acid substitutions. For both panels the elution fractions from each sample were normalized for GFP fluorescence and an equivalent amount of purified SufIss-GFP-His was loaded onto SDS/PAGE (4–15% Mini-PROTEAN TGX precast gradient gel) followed by Western blot using TatB and TatC mixed antibodies.