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. Author manuscript; available in PMC: 2021 Mar 25.
Published in final edited form as: J Am Chem Soc. 2020 Mar 11;142(12):5482–5486. doi: 10.1021/jacs.9b12185

Figure 2.

Figure 2.

The photocrosslinked adduct is MurJ×Lipid II. (a) Cells treated with the Lipid II biosynthesis inhibitor fosfomycin (fos) did not produce a crosslinked adduct. (b) Immunopurification of Flag-MurJF22pBpa with α-Flag resin prior to SDS-PAGE, biotinylation and immunoblotting confirmed that the 37 kD band contains MurJ. (c) When the Lipid II labeling enzyme PBP4 was omitted from the procedure shown in Figure 1c, the MurJ adduct was not detected. (d) When electroblots were pretreated with the D,D-carboxypeptidase DacA, which cleaves the terminal D-Ala residue from Lipid II, the MurJ adduct was not detected. See also Figures S1, S4–S9.