Expression and purification of regulatory domain of protein kinase C isoforms using bacterial expression system

초록

Protein kinase C (PKC) isoforms are a family of at least 10 serine/threonine protein kinases divided into three subfamilies: classical, novel & atypical, and are important modifiers of several cardiovascular phenomena, carcinogenesis & apoptosis, and are also promising therapeutic target for cancer and other indications. In our present study, we expressed and purified regulatory domain mutants of PKC isoforms. Bacterial protein expression plasmids were constructed by using pBADHisN3 vector. These expression vectors were transformed into TOP10 E.coli strain and expressed by Arabinose induction. Proteins were expressed as His tag protein and purified by Ni-NTA column. We checked the protein expression levels by SDS-PAGE analysis. We will use these proteins to characterize isoform-specific protein-protein interactions between PKC and other PKC-interacting proteins.