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. Author manuscript; available in PMC: 2017 Dec 1.
Published in final edited form as: Mol Microbiol. 2016 Sep 26;102(5):865–881. doi: 10.1111/mmi.13496

Figure 4.

Figure 4

Localization and lipidation of LtgA and LtgD. (A) Strains containing C-terminal 3XFLAG at the native site of ltgA or ltgD were fractionated into outer membrane (OM), total membrane (TM), and soluble (SOL) fractions using ultracentrifugation. Anti-FLAG antibodies were used to determine the subcellular localization of LtgA and LtgD, which localized to the same fractions as the OM control, PilQ. The strains used had a chromosomal cat gene to identify the cytoplasmic fraction and the presence of SecY was used as an inner membrane control. (B) The incorporation of the lipid [3H]-palmitate into recombinant LTs producing wild-type (WT), cysteine to alanine lipidation mutations (LM), or proteins lacking signal sequences (SM). The (*) indicates an active site mutant was used.