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. Author manuscript; available in PMC: 2018 Dec 4.
Published in final edited form as: Nat Chem Biol. 2018 Jun 4;14(7):730–737. doi: 10.1038/s41589-018-0061-0

Figure 5. Reconstitution of the lost adenylation activity of PtmA2 via site-directed mutagenesis.

Figure 5

a, Sequence alignment of PtmA2 and selected acyl-CoA synthetases. A3, A4, A8, and A10 motifs are shown with mutated residues highlighted with green boxes. See Supplementary Figure 10 for full sequence alignment. b,c, Total ion chromatograms (TICs; b) and extracted ion chromatograms (EICs; c) of PtmA2 mutant enzyme reactions. Shown are adenylation and complete (both adenylation and thioesterification) reactions with native and mutant PtmA2, and overlaid standards of 5, 6, and 7 (stds). See Supplementary Figures 14, 15, and 17 for reactions of other PtmA2 mutants, using free acid 8, and negative controls using boiled PtmA2. Chromatograms are representative examples of n>3 independent experiments.