Extended Data Fig. 4: ^PaMurA interacts with ^PaLpxC.

(a) H-^PaLpxC in vivo pulldowns. The expression status of H-^PaLpxC and F-^PaMurA before (input) and after (elution) co-affinity purification using Ni-NTA resin is indicated above the immunoblots. The variant of F-^PaMurA produced is indicated by WT for F-^PaMurA(WT) or * for F-^PaMurA(C117S). When indicated, 0.5 µg/mL CHIR-090 was added to cultures 1 hr prior to harvesting and was maintained in all lysis and wash buffers as detailed in the methods section. The following strains were used to generate the lysates: PA239 (lane 1), PA1013 (lane 2), PA1068 (lane 3), PA1071 (lanes 4 and 6), and PA1121 (lanes 5 and 7). Data are representative of at 3 biological replicates. (b) Purified F-^PaLpxC (2.5 µM) was mixed in a 1:1 ratio with purified H-MurA variants in the presence or absence of CHIR-090 (5.7 µM) as indicated. The mixtures were pulled down with anti-FLAG resin and the input and elution subjected to SDS-PAGE and Coomassie staining. Mobilities of H-MurA and F-LpxC are indicated by a cyan or gold carrot, respectively. Data are representative of 3 replicates. For gel source data, see Supplementary Fig. 1.