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. 2021 Jul 5;4:836. doi: 10.1038/s42003-021-02357-x

Fig. 5. Characterizing NorC conformational shifts in response to ICab and TPP+ binding in E. coli membranes.

Fig. 5

a Schematic showing the accessibility of C150 and T378 of NorC in different conformations in native membranes. FSEC profile with detergent purified NorC T378C incubated with ICab shows that the mutation does not alter NorC’s ability to interact with ICab. b Methoxypolyethylene glycol maleimide (or PEG5k Mal; avg mol wt = 5 kDa) accessibility assay in NorC’s vestibule in E. coli membranes. PEG5k Mal does not react with NorC WT in native membranes or in detergent purified form, while it readily interacts with NorC T378C, as seen in the western blot (probed with Anti-Histag antibody), but the reactivity decreases in presence of excess ICab, TPP+ or both. n = 8 for independent replicates. Source data are provided in the source data file.