Fig. 3: The NG domain of FlhF interacts with the cytoplasmic region of HubP.
From: Polar confinement of a macromolecular machine by an SRP-type GTPase
a Domain structure of the polar landmark protein HubP/FimV from S. putrefaciens and constructs used in this study. b Coomassie-stained SDS-PAGE employing StrepII-tagged HubP-C as bait and FlhF, its NG domain or the B-domain as prey. c Coomassie-stained SDS-PAGE employing StrepII-tagged HubP-C, StrepII-tagged HubP-C lacking the C-terminal FimV domain or a StrepII-tagged FimV domain alone as bait and NG-FlhF as prey. d Coomassie-stained SDS-PAGE probing the impact of GDP or GTP on interaction of StrepII-tagged HubP-C and NG-FlhF, acting as bait and prey, respectively. e GTPase activity of NG-FlhF in the presence of its stimulator FlhG, HubP-C and the two together. The bars are mean values from 5 independent experiments, each shown as dot, with the errors shown as standard deviation from the 5 independent experiments. f Coomassie-stained SDS-PAGE employing StrepII-tagged HubP-C as bait and FlhF alone and FlhF and FliG. For Fig. 3b, c, d, f: Each of the shown experiments were repeated in at least three biological replicates.
