2010•Protein and Peptide LettersRequires access
Production of Active MMP7 in E. coli and Its Application for Metalloproteinase Inhibitors Screening
Hiroshi Katsuno, Ryutaro Shirakawa, Kei Miyazaki, Yasuhiro Ozeki, Hidetaro Yasumitsu
Open publisher page 5 citations
Abstract
MMP-7 is the smallest metalloproteinase. Its unregulated activities and existence in serum are recently known to be tightly related with life-threatening disease such as cardiac disease and several cancers. The protein production is thought to be useful for its characterization and antibody generation. Although many attempts at bacterial expressions have been conducted, they were recovered as insoluble and inactive protein. In this study, after soluble expression, singlestep purification and conversion to active protease, it was applied for the screening secretory metalloproteinase inhibitors in conditioned media of human cancer cells. Keywords: MMP-7/matrilysin, matrix metalloproteinase, GST fusion, metalloproteinase inhibitor, soluble expression in E. coli, reverse-zymography