Production of active MMP7 in E. coli and its application for metalloproteinase inhibitors screening.
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, single-step purification and conversion to active protease, it was applied for the screening secretory metalloproteinase inhibitors in conditioned media of human cancer cells.
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.
H
H
Katsuno
R
R
Shirakawa
K
K
Miyazaki
Y
Y
Ozeki
H
H
Yasumitsu
D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005978:Glutathione / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020783:Matrix Metalloproteinase 7 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry / Q000378:metabolism
D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7; D005978:Glutathione