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Small phospho-donors phosphorylate MorR without inducing protein conformational changes.
Nathália S S Castro1, César A T Laia2, Biplab K Maiti1
1LAQV-REQUIMTE, Departamento de Química, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
Biophysical Chemistry
|June 9, 2018
Summary
This study shows bacterial response regulator MorR is phosphorylated by both acetylphosphate and phosphoramidate. Phosphorylation by phosphoramidate is more favorable and does not alter MorR protein structure.
Area of Science:
- Biochemistry
- Molecular Biology
- Microbiology
Background:
- Protein phosphorylation is a critical regulatory mechanism in biological systems.
- Two-component systems (TCS) are vital bacterial regulatory networks involving response regulators (RR) and kinase proteins.
- RR activity is modulated by phosphorylation, influencing cellular responses to environmental cues.
Purpose of the Study:
- To investigate the phosphorylation of MorR, a response regulator involved in molybdenum (Mo) homeostasis.
- To compare the phosphorylation efficiency of acetylphosphate and phosphoramidate on MorR in vitro.
- To analyze the structural impact of MorR phosphorylation.
Main Methods:
- In vitro phosphorylation assays using acetylphosphate and phosphoramidate as phospho-donors.
- Fluorescence spectroscopy to monitor changes in MorR upon phosphoramidate binding.
- Theoretical and computational modeling to assess phosphorylation favorability.
Main Results:
- MorR was successfully phosphorylated by both acetylphosphate and phosphoramidate.
- Phosphoramidate binding resulted in the quenching of MorR's tryptophan fluorescence.
- Computational analysis indicated that phosphoramidate is a more favorable phospho-donor for MorR compared to acetylphosphate.
Conclusions:
- Phosphorylated MorR exists as a monomer.
- The phosphorylation of MorR by acetylphosphate or phosphoramidate does not induce significant conformational changes in its protein structure.
- These findings provide insights into the regulatory mechanisms of bacterial TCS and Mo homeostasis.
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