Figure 3.

BpfD binds c-di-GMP via its degenerated EAL domain. Thermal shift assays (TSAs) were performed using the cytoplasmic part of BpfD (BpfD_S) (A), its GGDEF domain (B), or EAL (C) domain and c-di-GMP, cAMP and GTP. The various domains of BpfD (7.5 µM) were incubated in the presence of SYPRO Orange and various concentrations of the different compounds. The mix was then submitted to a temperature gradient from 20 to 70 °C. Graphs represent the first derivative of the fluorescence emission (-d(RFU)/dT, RFU: Raw Fluorescence Unit) as a function of temperature. The melting temperatures (Tm) of each protein are listed in the table (mean values with standard deviation, n = 2 to 6) (D). All graphs are representative of two independent experiments.