MW = 19.5 kDa. Recombinant matrix metalloproteinase-3 (MMP-3, stromelysin-1, transin) cloned from human cDNA, expressed in E.coli. The enzyme consists of the catalytic domain of human MMP-3, residues 105-265 (UniProtKB accession P08254). The enzyme has been inactivated by mutagenesis (E219A). This product is derived from MMP-3 Catalytic domain, and contain also the mutation F171D to increase its stability. No residual enzyme activity has been detected using the method described in the specific activity section of this data sheet.
100 110 120
M-F RTFPGIPKWR KTHLTYRIVN
130 140 150 160 170 180
YTPDLPKDAV DSAVEKALKV WEEVTPLTFS RLYEGEADIM ISFAVREHGD FYPFDGPGNV
190 200 210 220 230 240
LAHAYAPGPG INGDAHFDDD EQWTKDTTGT NLFLVAAHEI GHSLGLFHSA NTEALMYPLY
250 260 270
HSLTDLTRFR LSQDDINGIQ SLYGPPPDSP ET
> 95% by SDS-PAGE. The enzyme was observed as a single band migrating at a molecular weight of < 20 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).
Not detected. Activity described as U=100 pmol/min at 25°C using a colorimetric assay with thiopeptolide Ac-Pro-Leu- Gly-[2-mercapto-4-methyl-pentanoyl]-Leu-Gly-OC2
(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.
Johnson, L.L. et al. J. Biol. Chem. 275 (15), 11026-11033 (2000).
Chen, L. et al. J. Mol. Biol. 293 (3), 545-557 (1999).
Weingarten, H. & Feder, J. Anal. Biochem. 147 (2), 437-440 (1985).
Weingarten, H., Martin, R. & Feder, J. Biochemistry 24 (23), 6730-6734 (1985).