Purification and characterization of two inactive/latent protein phosphatases from pig brain.
Two inactive/latent protein phosphatases termed LP-1 (Mr 260,000) and LP-2 (Mr 350,000) were identified and purified from pig brain. Examination of molecular structures indicated that LP-1 has three subunits with molecular weights of 69,000, 55,000, and 34,000, respectively, whereas LP-2 contains only one subunit, with molecular weight of 49,000. When using phosphorylase a as a substrate, LP-1 was completely inactive and could be dramatically activated by freezing and thawing in 0.2 M 2-mercaptoethanol, whereas LP-2 contained some basal activity but could also be stimulated 40-fold by the same treatment. Kinetic analysis further indicated that both LP-1 and LP-2 enzymes dephosphorylate histone 2A, myelin basic protein, and phosphorylase a at a rather comparable rate, but the dephosphorylation of histone 2A and myelin basic protein seems to be spontaneously active. This, together with the results that trypsinolysis could specifically knock off phosphorylase phosphatase activity but caused no effect on the associated myelin basic protein/histone phosphatase activities, supports the notion that a two-site mechanism may possibly be involved in the regulation of substrate specificity of LP-1 and LP-2 enzymes in the central nervous system.
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000201:enzymology*; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification*; D011107:Polylysine; D013552:Swine; D014357:Trypsin / Q000378:metabolism
D046911:Macromolecular Substances; D011107:Polylysine; D010749:Phosphoprotein Phosphatases; D014357:Trypsin