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. 2021 May 3;118(19):e2101989118. doi: 10.1073/pnas.2101989118

Fig. 3.

Fig. 3.

LdtF modulates PG−Lpp linkages. (A) HPLC chromatograms of PG sacculi of WT carrying either vector (Ptrc::), pRB1(Ptrc::ldtF), pRB2 (Ptrc::ldtFH135A), or pRB3 (Ptrc::ldtFC143A). Cultures were grown to an A600 of ∼1 in LB containing 0.2 mM IPTG for PG isolation. The black arrow indicates the absence of tri-lys-arg peak. (B) Structures of muropeptides. (C) Mass spectra of peaks 3 and Y, showing molecular mass (M+H)+ of 1,155.58 and 1,795.78 Da. (D) Determination of PG−Lpp linkages in the WT and ldtF mutant was done by treating the intact sacculi with mutanolysin, followed by electrophoresis of the soluble muropeptides. Lpp-containing muropeptides were visualized by Western blot using anti-Lpp antibody. PG from the ldtABC mutant was used as negative control. Cell lysates of WT, ldtF, and ldtABC were used as controls (lanes 4, 5, and 6).