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Production of Active MMP7 in E. coli and Its Application for Metalloproteinase Inhibitors Screening

[ Vol. 17 , Issue. 5 ]

Author(s):

H. Katsuno, R. Shirakawa, K. Miyazaki, Y. Ozeki and H. Yasumitsu   Pages 568 - 572 ( 5 )

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

Affiliation:

Expert Laboratory for Life Environments (ELLE), Department of Environmental Biosciences, International Graduate School of Arts and Sciences, Yokohama City University, 22-2, Seto, Kanazawa-ku, Yokohama 236-0027, Japan.



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