Expression and purification of regulatory and catalytic domains of protein kinase C isoforms using maltose binding protein fusion expression system

초록

Protein kinase C (PKC) isoforms are a family of at least 10 serine/threonine kinases divided into three subfamilies: classical, novel & atypical, and are important modifiers of several cardiovascular phenomena, carcinogenesis & apoptosis and also promising therapeutic target for cancer and other indications. In our present study, we expressed and purified regulatory domain and catalytic domain mutants of PKC isoforms. Bacterial protein expression plasmids were constructed by using pMBPN3 vector. These expression vectors were transformed into BL21(DE3) E.coli strain and expressed by IPTG induction. Proteins were expressed as MBP fusion protein and purified by amylose resin. 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.