Fig 8.

In vitro analysis of the interaction of DgrA with c-di-GMP. Size exclusion chromatography was used to analyze the binding of purified DgrA to either c-di-GMP or GMP. (A to F) Absorbance of DgrA at 254 nm (A) and 280 nm (B) was monitored as a reference for comparison with the absorbance at 254 nm of individual nucleotides (C and D) or a mixture of DgrA and either c-di-GMP (E) or GMP (F). Elution of c-di-GMP at earlier retention volumes in the presence of DgrA is an indicator of complex formation. Equal quantities of 100 μM DgrA and 400 μM c-di-GMP or GMP were loaded in all experiments. Absorbance is shown in milli absorbance units (mAu).