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Description

CC1047 is a recombinant 452 amino acid polypeptide corresponding to human Pro-MMP-13, with an additional C-terminal His-tag with the sequence GVTHHHHHH expressed in E. coli and purified from periplasm. The calculated Mr of the recombinant protein is 51.681 kDa. Upon activation with APMA activated MMP-13 is formed., Matrix metalloproteinases (MMPs) are Zn2+- and Ca2+-dependent endopeptidases which function in the turnover of extracellular matrix components [Matrisian, 1992]. Main subfamilies of MMP are collagenases, gelatinases, stromelysins and membrane-type matrix metalloproteinases [Nagase, 1997]. Three homologous collagenases have been identified in human tissues: Interstitial collagenase, neutrophil collagenase and collagenase-3. These three enzymes cleave fibrillar collagens at a single site, generating fragments of approximately ¾ and ¼ the size of the original molecules. ProMMP-13 [Procollagenase-3] consists of 452 amino acids with a calculated Mr of 52.520 [Freije et al., 1994]. Due to N-linked glycosylation, the actual Mr is about 60000 Da [Knauper et al., 1996]. Within the protein the following domains and sequence regions can be distinguished [Freije et al., 1994, Knauper et al., 1996]: An N-terminal propeptide, which confers latency to the proenzyme, a Ca2+ and Zn2+-ion binding catalytic domain, a hinge region, and a C-terminal hemopexin-like domain. Latent procollagenase-3 can be activated by proteases such as stromelysin [ Knauper et al., 1996], gelatinase A, MT1-MMP and plasmin [ Knauper et al., 1996] or incubation with APMA Knauper et al., 1996]. The Mr of active collagenase-3 which begins with the N-terminal sequence YNVFPRTL is 48,000 Da. Collagenase-3 hydrolyzes type II collagen 5- to 6- times faster than type I and type III collagens. The enzyme also exhibits high activity towards gelatin and it degrades SERPINS as a1-antichymotrypsin and plasminogen activator inhibitor-2 [Knauper et al., 1996]. Collagenase-3 is inhibited in a 1:1 stoichiometric fashion by TIMP-1, TIMP-2 and TIMP-3. Collagenase-3 is expressed during fetal bone development [Stahlebackdahl, 1997]. In adult human tissues collagenase-3 has been detected only in pathological conditions: in malignant tumors [Freije et al., 1994], in chronic ulcers [Valaamo et al., 1997], in arthritic cartilage [Mitchell et al., 1996] and synovium [Wernicke et al., 1996].