Extended Data Fig. 6: Bacterial CARD domains mediate protein-protein interactions in multiprotein anti-phage defense complexes.

a, b, AlphaFold2-Multimer model of the trypsin-like protease and the NLR-like protein from the Lysobacter gasdermin system. The model confidence score for protein-protein interactions is depicted below. c, Predicted Aligned Error (PAE) of the AlphaFold2-Multimer predicted interactions between the trypsin-like protease and the NLR-like protein. d, AlphaFold2-Multimer models of the trypsin-like protease and the NLR-like protein from systems homologous to the Lysobacter gasdermin system. The model confidence score for protein-protein interactions is depicted below each model. Percent sequence identity between the Lysobacter NLR/protease and the respective protein is presented for each model. e, Western blot analyses of the experiments presented in Fig. 3h. The FLAG-tagged NLR-like protein was expressed together with either the full-length HA-tagged protease, the tagged protease in which the CARD domain was deleted or only with the HA-tagged CARD domain. Left panel, anti-FLAG beads were used to immunoprecipitate the FLAG-tagged NLR-like protein, and anti-HA antibody was used for western blotting. Right panel, anti-HA beads were used to immunoprecipitate the HA-tagged proteins, and anti-FLAG antibody used for western blotting. Representative of three replicates. f, Control experiments showing specificity of pulldown. Shown is SDS-PAGE with Coomassie stain analysis, with the following immunoprecipitation results: FLAG-tagged NLR-like protein from Lysobacter that was co-expressed with RFP; HA-tagged trypsin-like protease; HA-tagged CARD-deleted protease; and HA-tagged CARD domain from Lysobacter that was co-expressed with GFP g, Cell lysates of the samples used for immunoprecipitations shown in Fig. 3h, presented here as control. h, Cell lysates of the samples used for immunoprecipitations shown in panel f, presented here as control.