|Title||Purification of an oxidation-sensitive enzyme, pI258 arsenate reductase from Staphylococcus aureus.|
|Publication Type||Journal Article|
|Year of Publication||2003|
|Authors||Messens, J., J. C. Martins, I. Zegers, K. Van Belle, E. Brosens, and L. Wyns|
|Journal||J Chromatogr B Analyt Technol Biomed Life Sci|
|Date Published||2003 Jun 25|
|Keywords||Arsenite Transporting ATPases, Crystallography, X-Ray, Electrophoresis, Polyacrylamide Gel, Ion Pumps, Kinetics, Models, Molecular, Multienzyme Complexes, Nuclear Magnetic Resonance, Biomolecular, Oxidation-Reduction, Protein Conformation, Staphylococcus aureus|
Arsenate reductase (ArsC) from Staphylococcus aureus pI258 is extremely sensitive to oxidative inactivation. The presence of oxidized ArsC forms was not that critical for NMR, but kinetics and crystallization required an extra reversed-phase purification to increase sample homogeneity. The salt ions observed in the X-ray electron density of ArsC were investigated. Carbonate was found to have the lowest dissociation constant for activation (K(a)=1.1 mM) and potassium was stabilizing ArsC (DeltaT(m)=+6.2 degrees C). Also due to the use of these salt ions, the final yield of the purification had improved with a factor of four, i.e. 73 mg/l culture.
|Alternate Journal||J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.|
Purification of an oxidation-sensitive enzyme, pI258 arsenate reductase from Staphylococcus aureus.