CHARACTERIZATION OF THE OVINE-LENS PLASMA-MEMBRANE PROTEIN-KINASE SUBSTRATES

The cAMP-dependent protein-kinase-catalyzed phosphorylation of the two major intrinsic lens fiber cell plasma membrane proteins, MP20 and MP 26, is likely restricted to the inner cortical and nuclear regions of the lens in vivo. The ovine-lens-specific connexin, MP70. that has bee n identified as Cx50 in mice and Cx45.6 in the chick, is also a protei n kinase substrate although it does not appear to be phophorylated by a number of protein kinases including cAMP-dependent protein kinase, c almodulin-dependent protein kinase or protein kinase C. Rather, an ext rinsic lens membrane fraction was isolated which contained protein kin ase activity that catalyzed the phosphorylation of MP70: this protein kinase activity was cAMP-independent, Ca2+-independent, Mg2+-dependent . phosphorylated MP70 on a serine residue(s) and migrated with a molec ular mass of 35 kDa on a gel filtration column. Both MP70 phosphorylat ion and the endogenous protein kinase activity were restricted to the lens outer cortical region. This membrane-associated protein kinase ac tivity represents the first reported partial characterization of an en dogenous lens fiber cell protein kinase activity that catalyzes the ph osphorylation of a lens connexin protein. The phosphatase-induced shif t in the electrophoretic mobility of MP70 is not reversed by this prot ein kinase, indicating that MP70 is likely phosphorylated on different residues by two or more protein kinases.