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. 2018 Apr 19;2:1–13. doi: 10.1016/j.tcsw.2018.04.002

Fig. 6.

Fig. 6

BacA and PgpB increase lipid II consumption by PBP1B in the membrane. The activity of PBP1B was measured in proteoliposomes with PgpB, BacA or no phosphatase. (A) Radiolabelled lipid II was used as substrate and PG products were analysed by HPLC (as shown in Fig. 5A). The peak numbers relate to the muropeptide structures in Fig. 5D. (B) Quantification of the PG products obtained in two independent experiments (mean ± SD). (C) Quantification of the % peptides in cross-links obtained in two independent experiments (mean ± SD). (D) Using non-labelled lipid II and thin-layer chromatography allowed the detection of residual lipid II, C55-P and C55-PP. Control, proteoliposomes without enzyme with added lipid II and C55-P. (E) Quantification of lipid II and C55-P from 3 independent experiments as the one in panel D. The values are mean ± SD and asterisks indicate p-value comparisons with PBP1B (*p < 0.1 and **p < 0.05). HPLC assays were performed using E. coli polar lipids liposomes. For the TLC assay, artificial lipids were used plus LpoB(sol) was added at 4 µM.