Purification of the bifunctional enzyme, imidazoleglycerolphosphate dehydratase-histidinol phosphatase, of Salmonella typhimurium.
Mn2+ precipitation, antibody affinity chromatography, and selective proteolysis have been used to purify a bifunctional core of the hisB enzyme from Salmonella typhimurium. The core is homogeneous in subunit molecular weight; however, it is heterogeneous in its charge properties, probably as a result of multiple cleavage points produced by limited proteolytic digestion. The resistance of the enzyme to irreversible denaturation by urea allowed the use of urea to elute the hisB enzyme from a column of anti-hisB IgG immobilized on Sepharose. Heterogeneous oligomeric forms of the enzyme were demonstrated by electrophoretic analysis and exist as multiples of the 46,000 molecular weight monomer.
D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006642:Histidinol-Phosphatase / Q000302:isolation & purification*; D006836:Hydro-Lyases / Q000302:isolation & purification*; D007158:Immunologic Techniques; D046911:Macromolecular Substances; D008345:Manganese; D010447:Peptide Hydrolases; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification*; D012486:Salmonella typhimurium / Q000201:enzymology*; D014508:Urea
D046911:Macromolecular Substances; D008345:Manganese; D014508:Urea; D006642:Histidinol-Phosphatase; D010744:Phosphoric Monoester Hydrolases; D010447:Peptide Hydrolases; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase