C2 region-derived peptides inhibit translocation and function of beta protein kinase C in vivo.

RACK1 is a protein kinase C (PKC)-binding protein that fulfills the criteria previously established for a receptor for activated C-kinase (RACK). If binding of PKC to RACK anchors the activated enzyme near its protein substrates, then inhibition of this binding should inhibit translocation and function of the enzyme in vivo. Here, we have identified such inhibitors that mimic the RACK1-binding site on beta PKC. We first found that a C2-containing fragment, but not a C1-containing fragment of beta PKC, bound to RACK1 and inhibited subsequent beta PKC binding. The RACK1-binding site was further mapped; peptides beta C2-1 (beta PKC(209-216), beta C2-2 (beta PKC(186-198)), and beta C2-4 (beta PKC(218-226), but not a number of control peptides, bound to RACK1 and inhibited the C2 fragment binding to RACK1. Peptides beta C2-1, beta C2-2, and beta C2-4 specifically inhibited phorbol ester-induced translocation of the C2-containing isozymes in cardiac myocytes and insulin-induced beta PKC translocation and function in Xenopus oocytes. Therefore, peptides corresponding to amino acids 186-198, and 209-226 within the C2 region of the beta PKC are specific inhibitors for functions mediated by beta PKC.