Subject(s)
Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Escherichia coli/enzymology , Mannitol/metabolism , Phosphoenolpyruvate Sugar Phosphotransferase System/chemistry , Phosphoenolpyruvate Sugar Phosphotransferase System/metabolism , Carbon Isotopes , Nitrogen Isotopes , Nuclear Magnetic Resonance, Biomolecular , Phosphorylation , Protons , Substrate SpecificityABSTRACT
Residual dipolar couplings between 15N and 1H nuclear spins in HPr were used to determine the protein's orientation in a medium of bicelles, oriented by a magnetic field. In the case of wild-type HPr the protein's non-spherical shape can explain its orientation in this medium. In the case of the F48W mutant it was found that at least one other mechanism contributes to the observed orientation of the protein, to a degree that depends on the concentration of phosphate ions in the medium. We propose that the F48W mutant has a weak affinity towards the bicelle-surfaces that decreases with increasing phosphate concentrations. We used an order-parameter description to analyse this situation and to determine the axis of main order and the sign of the order parameter pertaining to this additional orientation mechanism.
