Fig. 2: Gp54Bas11 binds directly to the antitoxin region of CapRelSJ46.
From: A bacterial immunity protein directly senses two disparate phage proteins
a, Schematic of the domain organization of CapRelSJ46 (top) and cartoon representation of the crystal structure of CapRelSJ46 coloured by domains (bottom). Active site G-loop Y155 and ATP-coordination residues R79 and R116 of the toxin domain (toxSYNTH) are highlighted in red. b, Flag-tagged CapRelSJ46 (CapRelSJ46–Flag) or chimera–Flag was immunoprecipitated from cells producing CapRelSJ46–Flag or chimera–Flag and haemagglutinin (HA)-tagged Gp54Bas11 (wild type or the G24D variant), and probed for the presence of the indicated Gp54Bas11 variant using the HA tag. Lysates used as input for immunoprecipitation (IP) were probed as controls for expression levels. Image shown is representative of two biological replicates. c, Binding of CapRelSJ46 to the wild-type or G24D variant of Gp54Bas11 monitored by ITC, with binding affinity (Kd) and stoichiometry (N) noted. d, Left, schematic of the CapRel constructs. Right, serial dilutions of the Bas11 phage spotted on lawns of cells harbouring either the indicated CapRel constructs or an empty vector. e, Serial dilutions of cells producing CapRelSJ46 or the chimera from its native promoter and wild-type Gp54Bas11 from an arabinose-inducible promoter on medium containing glucose or arabinose.
