Chemical Synthesis of a PKC C1b Domain by a Peptide Ligation Method and Expression of the Protein in E. coli and Their Application

摘要

Protein kinase C, PKC, is a family of enzymes for phosphorylation, which is specific for Ser and Thr residues. PKC family comprises at least 11 isozymes, which play fundamental roles in signaling pathways that regulate cell cycle progression, differentiation, and apoptosis. PKC activity is regulated by phosphorylation, by calcium, and by association with phospholipids and a physiological ligand, diacylglycerol (DAG). PKC has also been proven to be involved in problematic diseases, such as cancer and Alzheimer's diseases. Thus, it has been established as an important therapeutic target. Marquez's group has developed conformationally constrained analogs of DAG based on gamma-lactone templates [1,2]. In this study, we synthesized a PKCδ C1b domain, which comprises a binding pocket of DAG, by a native chemical ligation method. This domain was also expressed in E. coli to study structural analysis of its complex with the above DAG analogs.^