Matrix metalloproteinases (MMPs) are Zn2+-and Ca2+-dependent endopeptidases which function in the turnover of extracellular matrix components (1). Presently, eighteen secreted MMPs and five membrane-type MMPs (2–5) are known to be expressed in vertebrates. Human MT1-MMP consists of 559 amino acid residues with a calculated Mr of 63516 (2,3). The following domains and sequence regions are distinguished in MT1-MMP: Prodomain (Ser1-Arg88), catalytic domain (Tyr89-Gly261), junction between catalytic domain and hemopexin domain (Gly262-Gly292), hemopexin-like domain (Pro293-Cys485) and C-terminal sequence (Pro486-Val559) with transmembrane segment. When expressed in cells, the catalytic domain and the hemopexin domain of MT1-MMP become exposed on cell surfaces (2,6). A soluble form of MT1-MMP without transmembrane segment has been found in culture medium of a breast carcinoma cell line (7). MT1-MMP is expressed in adult lung, placenta, kidney, ovaries, intesting, prostate and spleen (3). Increased amounts of the enzyme are found in tumor tissues such as lung carcinoma (2), gastric carcinoma (8), breast, head and neck carcinoma (9). MT1-MMP is activated by removal of its prodomain. The reaction is catalyzed by furin, a subtilysin-type serine protease, which recognizes a motif of four basic amino acid residues located between the prodomain and catalytic domain (6). MT1-MMP activates progelatinase A (2,10,11) and procollagenase-3 (12) by proteolytic cleavage of their domains. The ability of MT1-MMP to activate other matrix metalloproteinases provides potential for enhanced pericellular proteolysis in physiological and pathological processes. In particular, activation of progelatinase A by MT1-MMP is considered to contribute to local degradation of extracellular matrix during cell migration and proliferation. MT1-MMP also hydrolyzes fibrillar collagens I, II and III into characteristic 3/4 and 1/4 fragments (13) and it cleaves a number of other ECM proteins, including fibronectin, vitronectin, laminin-1 and dermatan sulfate proteoglycan (6,13). The activity of MT1-MMP is poorly inhibited by TIMP-1 but efficiently inhibited by TIMP-2 and TIMP-3 (11). The recombinant prodomain and catalytic domain of MT1-MMP is useful for the study of Progelatinase A (MMP-2) activation and the degradation of proteins of the extracellular matrix. The enzyme allows screening of matrix metalloproteinase inhibitors and characterization of inhibitor action.
Source:Recombinant polypeptide sequence produced as a periplasmic protein in E. coli. The pro-MT1-MMP deletion mutant consists of MT1-MMP residues corresponding to Ser1-Gly265 followed by the His-tag sequence Leu-Val-Thr-(His)6. The calculated Mr of the recombinant proenzyme is 30860 Da. Upon activation with trypsin the prodomain sequence Ser1-Arg88 is removed from recombinant pro-MT1MMP and enzymatically active catalytic domain beginning with Tyr89 is liberated.
Applications:Useful as an antigen standard in immunoassays. The proenzyme can be activated with trace amounts of MT1-MMP catalytic domain (1,2).
Storage and Stability:May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
仕様
Size：5ug
Source Antigen：Recombinant, E. coli
Grade：Purified
Purity：Appears as a predominant band at 58kD in SDS-PAGE (> 80% total protein)
Form：Provided as a liquid in 50mM Tris-HCl, pH 7.5, 150mM sodium chloride, 5mM CaCl2.
