Purification and properties of rabbit liver phosphorylase phosphatase.
A procedure for the purification of rabbit liver phosphorylase phosphatase is described. The specific activity of the preparation is 2,100 units/mg of protein, representing a 25,000-fold purification. During the initial steps of the purification a large activation of enzyme activity was observed. The molecular weight of the purified enzyme was estimated by Sephadex G-75 chromatography to be 35,000, and by sucrose density ultracentrifugation to be 34,000 (2.9 S). On Na dodecyl-SO4 polyacrylamide disc gel electrophoresis a single component with a molecular weight of 34,000 was observed. The pH optimum is 6.9 to 7.4, and the Km for rabbit muscle phosphorylase alpha is 2 muM. The same procedure is also applicable to the extensive purification of phosphorylase phosphatase from rabbit muscle.
D000818:Animals; D005947:Glucose / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010765:Phosphorylase Phosphatase / Q000302:isolation & purification / Q000378:metabolism*; D011817:Rabbits; D013806:Theophylline / Q000494:pharmacology
D013806:Theophylline; D010765:Phosphorylase Phosphatase; D010744:Phosphoric Monoester Hydrolases; D005947:Glucose