Purification and partial characterization of a unique group of phosphoproteins from rat milk whey

Abstract

A group of four electrophoretically distinct but related proteins has been isolated from rat milk whey. These have been classified as P.1 to P.4 in order of their decreasing electrophoretic mobility in an alkaline buffer system. All four members of this group of proteins are immunologically identical apparently constitute a single protein which is differentially phosphorylated in the ratio of 3:2:1:0. Sodium dodecyl sulfate gel electrophoresis yielded constant estimates of molecular weight of approximately 20,700. Enzymatic dephosphorylation of purified P.1 generated the other three members of the group, demonstrating the commonality of the peptide component. This group of proteins in total makes a significant contribution of the total whey proteins being approximately 5 mg/ml in whole rat milk in the approximate proportion of 1:1.5:2:1.5. The P.1, P.2, and P.3 were isolated in sufficient quantities to permit further characterization. The partial amino acid analyses of each of the three forms were similar. They had commonextinction coefficients, E1(1)% cm A280, of 4.9 and A280/A290 ratio of about 1.36.