Overexpression of recombinant human procathepsin B in E. coli

Abstract

Cathepsin B is a lysosomal cysteine proteinase that is implicated in the processes of tumor invasion, metastasis, and Alzheimer's disease. Development of specific inhibitors of cathepsin B could lead to the discovery of therapeutic agents for treatment of many types of carcinomas, as well as Alzheimer's disease. The objective of this research is to produce functional recombinant cathepsin B enzyme which will be used in screening of new cathepsin B inhibitors. Human procathepsin B cDNA has been successfully amplified by PCR and sub‐cloned into pET15b expression vector. Overexpression of the procathepsin B protein has been accomplished by IPTG induction. The ability to produce large quantities of procathepsin B enzyme will permit a drug discovery program for development of newer effective inhibitors of cathepsin B.Supported by National Cancer Institute at NIH (Grant No. 3R15CA086933‐04 and 3R15CA086933‐04A2S1)