MW = 17.5 kDa calculated. Matrix Metalloproteinase-20 (MMP-20, Enamelysin, Enamel metalloproteinase) catalytic domain cloned from human cDNA, expressed in E. coli. The enzyme consists of the catalytic domain of human MMP-20 (residues 113-271). Residues numbers are based on the unprocessed precursor). UniProtKB accession O60882.
120 130 140 150 160 M-GEPKWKKN TLTYRISKYT PSMSSVEVDK AVEMALQAWS SAVPLSFVRI 170 180 190 200 210 NSGEADIMIS FENGDHGDSY PFDGPRGTLA HAFAPGEGLG GDTHFDNAEK 220 230 240 250 260 WTMGTNGFNL FTVAAHEFGH ALGLAHSTDP SALMYPTYKY KNPYGFHLPK 270 DDVKGIQALY 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.
0.1 mg/mL solution in Tris 20 mM pH 7.2, CaCl2 10 mM, ZnCl2 0.1 mM, NaCl 0.3 M, acetohydroxamic acid (AHA) 0.5 M. The concentration is calculated by the analysis of the absorbance at 280 nm (ε280 = 28420 M-1cm-1 calculated).
> 50 U/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.
-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.
Yamakoshi, Y., et al. J Biol Chem. 2006 Dec 15;281(50):38235-43. Epub 2006 Oct 17.