MW = 19.3 kDa calculated. Recombinant Matrix Metalloproteinase-7 (MMP-7, Matrilysin, Uterine metalloproteinase) cloned from human cDNA, expressed in E. coli. The enzyme consists of the catalytic domain of human MMP-7 (residues 95-267, UniProtKB accession P09237).
100 110 120 130 140 M-YSLFPN SPKWTSKVVT YRIVSYTRDL PHITVDRLVS KALNMWGKEI 150 160 170 180 190 PLHFRKVVWG TADIMIGFAR GAHGDSYPFD GPGNTLAHAF APGTGLGGDA 200 210 220 230 240 HFDEDERWTD GSSLGINFLY AATHELGHSL GMGHSSDPNA VMYPTYGNGD 250 260 PQNFKLSQDD IKGIQKLYGK RSNSRKK
> 95% by SDS-PAGE. The protein is observed, in denaturing conditions, as a single band migrating at a molecular weight between 18.8 kDa and 25.0 kDa.
0.2 mg/mL solution in Tris 20 mM pH 7.2, CaCl2 10 mM, ZnCl2 0.1 mM, NaCl 0.3 M, acetohydroxamic acid (AHA) 0.5 M. The concentration is calculated by the analysis of the absorbance at 280 nm (ε280 = 33920 M-1cm-1 calculated).
> 8U/μg. Activity described as U=100 pmol/min at 25°C using a colorimetric assay with thiopeptide Ac-Pro-Leu-Gly-[2- mercapto-4-methyl-pentanoyl]-Leu-Gly-OC2H5 (Biomol) as substrate.
-80°C. After initial defrost, aliquot the product into individual tubes and refreeze at -80°C.
Avoid repeated freeze/thaw cycles.
Enzyme kinetic studies, cleavage of target substrates and screening of inhibitors.
McCawley, L.J. & Matrisian, L.M. Curr. Opin. Cell Biol. 13 (5), 534-540 (2001).
Stetler-Stevenson, W.G. & Yu, A.E. Semin. Cancer Biol. 11 (2), 143-152 (2001).
Browner, M.F., Smith, W.W. & Castelhano, A.L. Biochemistry 34 (20), 6602-6610 (1995).