||Recombinant human Metalloproteinase-9 (MMP-9, 92 kDa type IV collagenase, 92 kDa gelatinase, Gelatinase B, GELB) catalytic domain with fibronectins domains cloned from human cDNA, was expressed inE. coli.The recombinant enzyme consists of the catalytic domain of human MMP-9 (residues 112-445) with a N-term T7 tag. MW= 37.7kDa.
||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 MMPs are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The enzyme encoded by this gene degrades type IV and V collagens. Studies in rhesus monkeys suggest that the enzyme is involved in IL-8-induced mobilization of hematopoietic progenitor cells from bone marrow, and murine studies suggest a role in tumor-associated tissue remodeling.
||> 95% by SDS-PAGE. In an SDS-PAGE gel, the enzyme runs as a single band migrating between 35 and 45 kDa.
||0.15mg/ml in 50mM Tris pH 7.2, 5mM CaCl2, 0.1mM ZnCl2, 0.3M NaCl, 0.5M Acetohydroxamic Acid (AHA), 10% glycerol, Brij35 0.05%. The concentration is calculated from the absorbance at 280nm (ε280 = 67100 M-1 cm-1calculated).
||>10U/μg. 1U=100pmol/min at 25ºC using a colorimetric assay with thiopeptolide Ac-Pro-Leu-Gly-[2-mercapto-4-methyl-pentanoyl]-Leu-Gly-OC2H5 (Biomol).
||Study enzyme kinetics, cleavage of target substrates and screen for inhibitors.
||-80℃. It is recommended that thawing and dilution of the enzyme be done in ice and within as short a time as possible before start of the assay. After initial defrost, aliquot product into individual tubes and refreeze at -80℃. Avoid repeated freeze/defrost cycles.