Small phospho-donors phosphorylate MorR without inducing protein conformational changes

Phosphorylation is an essential mechanism of protein control and plays an important role in biology. The two-component system (TCS) is a bacterial regulation mechanism mediated by a response regulator (RR) protein and a kinase protein, which synchronize the regulatory circuit according to the environment. Phosphorylation is a key element in TCS function as it controls RR activity. In the present study, we characterize the behavior of MorR, an RR associated with Mo homeostasis, upon acetylphosphate and phosphoramidate treatment in vitro. Our results show that MorR was phosphorylated by both phospho-donors. Fluorescence experiments showed that MorR tryptophan emission is quenched by phosphoramidate. Furthermore, theoretical and computational results demonstrate that phosphorylation by phosphoramidate is more favorable than that by acetylphosphate. In conclusion, phosphorylated MorR is a monomeric protein and phosphorylation does not appear to induce observable conformational changes in the protein structure. [Display omitted] •Phosphoramidate and acetylphosphate phosphorylate MorR in several aminoacid residues•Phosphorylation does not induce MorR oligomerization.•MorR-Tryptophan emission is quenched in the presence of phosphoramidate.•Phosphoramidate is energetic favorable to phosphorylation.