EDTA-extractable proteins from calf lens fiber membranes are phosphorylated by Ca2+-phospholipid-dependent protein kinase.

A distinct group of EDTA-extractable proteins (EEP), being a major protein component of calf lens fiber membranes, is bound to these membranes in a calcium-dependent way. Both purified and membrane-bound EEP can be phosphorylated in vitro by a Ca2+-activated, phospholipid-dependent protein kinase (protein kinase C). In general, this protein kinase preferentially phosphorylates serine and threonine residues of protein substrates. Phosphoamino-acid analysis of the two major bands of EEP phosphorylated by protein kinase C, representing the 33,000 + 34,000 EEP proteins and the 30,700-31,800 proteins, respectively, revealed differences in the phosphoamino-acid patterns. For the 33,000 + 34,000 EEP proteins, only phosphothreonine was detected whereas for the 30,700-31,800 proteins, the label was incorporated in both threonine and serine residues. No label was found on tyrosine residues. These results implicate differences in the primary structure of the individual EEP proteins. Regarding previous observations that EEP is a main protein component of lens fiber junctions and of the many covering epithelial and endothelial cells, and considering the fact that protein kinase C is involved in cell-cell communication, growth and differentiation processes we suggest that a correlation exists between phosphorylation-dephosphorylation of EEP and the regulation of a number of cellular processes.