Phosphorylation of caseins, present evidence for an amino acid triplet code posttranslationally recognized by specific kinases

Abstract

The fifty of so phosphorylated hydroxyamino acid residues hitherto investigated in caseins from different species have been found to occur in tripeptide sequences -Ser/Thr-X-A- where X represents any amino acid residue and A is an acidic residue. This strongly suggests that phosphorylation of caseins involves basically the stepwise enzymatic recognition of primary and secondary anionic amino acid triplets where the determinants are dicarboxylic residues and phosphoseryl residues, respectively. Studies of genetic variants of bovine caseins have provided clear-cut evidence for the actual occurrence of the former recognition sites. The occurrence of the above tripeptide sequences is a necessary but not a sufficient condition for phosphorylation of caseins to occur. Possible factors of constraint such as different intrinsic properties of both phosphate acceptor residues and acidic determinants, the characteristics of the local environment in terms of overall charge and hydrophilicity, secondary structure and steric hindrance, an insufficient available pool of casein kinase(s)... are discussed. All evidence now available supports the concept that phosphorylation of caseins is a posttranslational event and it is suggested that the process may occur during the transfer of completed polypeptide chains from the smooth endoplasmic reticulum to the Golgi apparatus where most of phosphate incorporation is presumably carried out. This organelle is rich in membrane-bound specific cyclic AMP-independent kinase(s) able in vitro to rephosphorylate specifically although not completely phosphatase-treated caseins and caseinophosphopeptides.