title: Purification of matrix metalloproteinases by column chromatography.
abstract: Matrix metalloproteinases (MMPs) are zinc endopeptidases composed of 23 members in humans, which belong to a subfamily of the metzincin superfamily. They play important roles in many pathophysiological events including development, organogenesis, angiogenesis, tissue remodeling and destruction, and cancer cell proliferation and progression by degradation of extracellular matrix (ECM) and non-ECM proteins and interaction with various molecules. Here, we present standard protocols for purification of native proMMPs (proMMP-1, -2, -3, -7, -9 and -10) and recombinant MT1-MMP (MMP-14) using conventional column chromatography. Purification steps comprise the initial common step [diethylaminoethyl (DEAE)-cellulose, Green A Dyematrex gel and gelatin-Sepharose columns], the second step for removal of nontarget proMMPs by immunoaffinity columns (anti-MMP-1 and/or anti-MMP-3 IgG-Sepharose columns) and the final step for further purification (IgG-Sepharose, DEAE-cellulose, Zn2+-chelate-Sepharose and/or gel filtration columns). Purified proMMPs and MMP are functionally active and suitable for biochemical analyses. The basic protocol for the purification from culture media takes approximately 7-10 d.
authors.affiliation: Department of Biochemistry, Nippon Dental University, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.
authors.forename: Kazushi
authors.initials: K
authors.lastname: Imai
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Okada
mesh_terms: D002845:Chromatography / Q000379:methods*; D003636:DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D020782:Matrix Metalloproteinases / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D003636:DEAE-Cellulose; D020782:Matrix Metalloproteinases