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. Author manuscript; available in PMC: 2019 Sep 15.
Published in final edited form as: Mol Microbiol. 2018 Sep 15;109(5):633–641. doi: 10.1111/mmi.14002

Figure 3. Support of PG lipid-II flipping in vivo by a WzxC variant.

Figure 3

Cells of CAM290 [murJ(A29C)] harboring plasmid pCS124 [Plac::wzxC(WT)-flag] or pDF2 [Plac::wzxC(V252M)-flag] were grown in labeling medium to an OD600 of 0.2. [3H]-mDAP was then added to radiolabel PG precursors. After a 15 min labeling period, MTSES and ColM were added to block MurJ(A29C) activity and cleave flipped lipid II, respectively. Just prior to cell lysis, cells were collected by centrifugation and fractionated to measure radioactivity in the PG lipid precursor pool (A) and soluble ColM cleavage product (B). WT and Mut denote WzxC(WT) and WzxC(V252M), respectively. The means and the standard error of means (SEMs) from three experiments are shown. P-values were calculated with two-tailed unpaired Student’s t-test. *, p < 0.05; **, p < 0.01; n.s., not significant. cpm = counts per minute.