||Recombinant Human matrix metalloproteinase-14 (MMP-14, Membrane-Type Matrix Metalloproteinase1, MT1-MMP) cloned from human cDNA was expressed inE.coli. The enzyme consists of the catalytic domain of human MMP-14 (residues 114-290 swissprot accession P50281) with a C-term purification tag. The protein has been mutated to increase the stability. The catalytic activity rates are not affected by the mutation. The C-terminal tag is cleaved during purification. MW=21.3kDa.
||Matrix metalloproteinase-14 is an enzyme that in humans is encoded by the MMP14 gene. Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP"s are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. However, the protein encoded by this gene is a member of the membrane-type MMP (MT-MMP) subfamily; each member of this subfamily contains a potential transmembrane domain suggesting that these proteins are expressed at the cell surface rather than secreted. This protein activates MMP2 protein, and this activity may be involved in tumor invasion.
||> 95% by SDS-PAGE. The enzyme was observed as a single band migrating at a molecular weight of > 20kDa.
||>150U/μg. Activity described as U=100pmol/min at 25ºC using a colorimetric assay with thipeptolide 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.
||0.2mg/ml in 20mM Tris, pH7.2, 10mM CaCl2, 0.1mM ZnCl2, 0.3M NaCl, 0.5M Acetohydroxamic Acid (AHA). The concentration is calculated from the absorbance at 280nm (e280=33000M-1 cm-1).
||Under the above described conditions, to avoid precipitation or protein self digestion, the product can be concentrated to a maximum of 200μM.
||-80ºC. The enzyme is stable at -20ºC for at least 1 week. After initial defrost, aliquot enzyme into individual tubes and refreeze at -80ºC. Avoid repeated freeze/defrost cycles.