||Recombinant Human matrix metalloproteinase-1 (MMP-1, fibroblast collagenase, collagenase-1, interstitial collagenase) cloned from human cDNA was expressed inE. coli. The enzyme consists of the catalytic domain of human MMP-1 (residues 106-261, swissprot accession P03956). MW=17.6 kDa.
||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. This gene encodes a secreted enzyme which breaks down the interstitial collagens, types I, II, and III. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.In addition, mechanical force may increase the expression of MMP1 in human periodontal ligament cells.
||> 95% by SDS-PAGE. The enzyme was observed as a single band migrating at a molecular weight <20 kDa.
||>100U/μg. Activity described as U=100 pmol/min at 25ºC 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.
||0.2mg/ml in 20mM Tris, pH 7.2, 10mM CaCl2, 0.1mM ZnCl2, 0.3M NaCl, 0.2M Acetohydroxamic Acid (AHA). The concentration is calculated from the absorbance at 280nm. (e280 = 25440 M-1 cm-1).
||Under the above described conditions, to avoid precipitation or protein dimerization, the product can be concentrated to a maximum concentration of 1mM.
||-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.