| Abstract: | The activating factor FA of the ATP.Mg-dependent protein phosphatase FcM was purified to near homogeneity from pig brain by a procedure involving chromatography on phosphocellulose, phosvitin-Sepharose 4B, and Blue Sepharose CL-6B. A specific myelin basic protein (MBP) kinase was found to co-purify with FA in a constant ratio throughout purification. It also proved impossible to separate the two activities on nondenaturing gel electrophoresis and 5-20% sucrose density gradient ultracentrifugation. Kinetic study indicated that MBP, presumably a substrate for FA, could compete with FcM for FA and thereby prevent the FA-mediated activation of the FcM activity. All the results taken together demonstrate that MBP kinase and FA are localized on the same protein. This, together with the data that FA, by activating the ATP.Mg-dependent phosphatase, promotes the dephosphorylation of 32P]MBP, phosphorylated by FA itself, suggests the evidence for a protein bearing two opposing activities involved in the regulation of brain functions. Moreover, since FA is tightly associated with the purified brain myelin membrane, the results further support the notion that FA may well be an endogenous protein kinase responsible for the cyclic phosphorylation-dephosphorylation of the central nervous system myelin. |
