MW = 17.6 kDa calculated. Recombinant matrix metalloproteinase-1 (MMP-1, fibroblast collagenase, collagenase-1, interstitial collagenase) cloned from human cDNA, expressed in E. coli. The enzyme consists of the catalytic domain of human MMP-1 (residues 106-261, UniProtKB accession P03956).
110 120 130 140 150 M-NPRWE QTHLTYRIEN YTPDLPRADV DHAIEKAFQL WSNVTPLTFT 160 170 180 190 200 KVSEGQADIM ISFVRGDHRD NSPFDGPGGN LAHAFQPGPG IGGDAHFDED 210 220 230 240 250 ERWTNNFREY NLHRVAAHEL GHSLGLSHST DIGALMYPSY TFSGDVQLAQ 260 DDIDGIQAIY G
> 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= 28420 M-1cm-1 calculated).
-80°C. After initial defrost, aliquot the product into individual tubes and refreeze at -80°C.
Avoid repeated freeze/thaw cycles.
> 100U/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.
Enzyme kinetic studies, cleavage of target substrates and screening of inhibitors.