Purification of a soluble monoisozyme of nucleoside diphosphate kinase from chick brain: exploitation of ionic characteristics.
A procedure for the rapid purification of nucleoside diphosphate kinase, 24 h with a single operator, from the chick brain soluble fraction is described. The influence of the ionic conditions on the association-disassociation properties of the enzyme are exploited to obtain yields of 30% from the crude homogenate. The enzyme has been purified 500-fold with a maximal specific activity of 1500 mumol/min/mg at 25 degrees C (using thymidine diphosphate as the phosphate acceptor and ATP as the donor) and is demonstrated to be monoisozymic.
D000645:Ammonium Sulfate; D000818:Animals; D001923:Brain Chemistry; D002645:Chickens; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D007527:Isoenzymes / Q000302:isolation & purification*; D008722:Methods; D009701:Nucleoside-Diphosphate Kinase / Q000302:isolation & purification*; D010770:Phosphotransferases / Q000302:isolation & purification*
D007527:Isoenzymes; D010770:Phosphotransferases; D009701:Nucleoside-Diphosphate Kinase; D000645:Ammonium Sulfate