MW = 17.6 kDa calculated. Matrix Metalloproteinase-19 isoform 1 RASI-1 (MMP-19, Matrix Metalloproteinase RASI, MMP-18) catalytic domain cloned from human cDNA, expressed in E. coli. The enzyme consists of the catalytic domain of human MMP-19, (residues 101-256, UniProtKB accession Q99542).
110 120 130 140 M- LGRWRKKHLT FRILNLPSTL PPHTARAALR QAFQDWSNVA 150 160 170 180 190 PLTFQEVQAG AADIRLSFHG RQSSYCSNTF DGPGRVLAHA DIPELGSVHF 200 210 220 230 240 DEDEFWTEGT YRGVNLRIIA AHEVGHALGL GHSRYSQALM APVYEGYRPH 250 FKLHPDDVAG IQALYG
> 95% by SDS-PAGE. The protein is observed, in denaturing conditions, as a single band migrating at a molecular weight between 14.4 and 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) 0.2 M. The concentration is calculated by the analysis of the absorbance at 280 nm (ε280 = 25440 M-1cm-1 calculated).
> 1.5 U/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.
Linksvan Horssen J., et al., Neuropathol Appl Neurobiol, 2006, 32(6):585-93.