MW = 17.9 kDa calculated. Matrix Metalloproteinase-2 (MMP-2, Gelatinase A, Type IV collagenase) catalytic domain without fibronectin domains cloned from human cDNA, expressed in E. coli. The enzyme consists of the catalytic domain of human MMP-2 fibronectin deficient (residues 115-214 and 393-447 where the residue numbers are based on the unprocessed precursor, UniProtKB accession P08253).
120 130 140 150 M-RKPKWD KNQITYRIIG YTPDLDPETV DDAFARAFQV 160 170 180 190 200 WSDVTPLRFS RIHDGEADIM INFGRWEHGD GYPFDGKDGL LAHAFAPGTG 210 VGGDSHFDDD ELWT 400 QGYSLFLV 410 420 430 440 450 AAHEFGHAMG LEHSQDPGAL MAPIYTYTKN FRLSQDDIKG IQELYGASPD
> 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.
0.15 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, glycerol 10%, Brij-35 0.05%. The concentration is calculated by the analysis of the absorbance at 280 nm (ε280= 32430 M-1cm-1 calculated).
> 40U/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.
Steffensen B. at al. J Biol Chem. 1995 May 12; 270(19):11555-66
Banyai L. et al. FEBS Lett. 1991 Apr 22; 282(1):23-5.