MW = 18.6 kDa. Recombinant matrix metalloproteinase-13 (MMP-13, collagenase-3, col3) cloned from human cDNA, expressed in E. coli. The enzyme consists of the catalytic domain of human MMP-13 (residues 104-268, UniProtKB accession P45452). The enzyme has been inactivated by mutagenesis (E223A). This product is derived from MMP-13 Catalytic domain and contain also the mutation F175D to increase its stability. No residual enzyme activity has been detected using the method described in the specific activity section of this data sheet.
110 120 130 140 150 YNVFPRT LKWSKMNLTY RIVNYTPDMT HSEVEKAFKK AFKVWSDVTP 160 170 180 190 200 LNFTRLHDGI ADIMISFGIK EHGDDYPFDG PSGLLAHAFP PGPNYGGDAH 210 220 230 240 250 FDDDETWTSS SKGYNLFLVA AHEFGHSLGL DHSKDPGALM FPIYTYTGKS 260 HFMLPDDDVQ GIQSLYGP
> 95% by SDS-PAGE. The protein is observed, in denaturing conditions, as a single band migrating at a molecular weight of about 18.4 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 = 29910 M-1cm-1 calculated).
Not detected. 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.
F.J.Moy et al. J Mol Biol. 2000 Sep 22; 302(3):671-89.
S. Cowell et al. Biochem. J. 1998, 331, 453.
G. Murphy and V. Knäuper Matrix Biol. 1997, 15, 51.