MMP-14, human, prodomain, catalytic domain, and hemopexin domain, E. coli recombinant

BACKGROUND: Matrix metalloproteinases (MMPs) are Zn2+- and Ca2+-dependent endopeptidases which function in the turnover of extracellular matrix components [Matrisian, 1992]. Presently, eighteen secreted MMPs and five membrane-type MMPs [Sato et al., 1994; Will & Hinzmann, 1995; Takino et al., 1995; Puente et al., 1996] are known to be expressed in vertebrates. Human MT1-MMP consists of 559 amino acid residues with a calculated Mr of 63516 [Sato et al., 1994; Will & Hinzmann, 1995]. 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. A soluble form of MT1-MMP without transmembrane segment has been found in culture medium of a breast carcinoma cell line [Imai et al., 1996].

MT1-MMP is expressed in adult lung, placenta, kidney, ovaries, intestine, prostate and spleen [Will & Hinzmann, 1995]. Increased amounts of the enzyme are found in tumor tissues such as lung carcinoma [Butler et al., 1998], gastric carcinoma [Nomura et al., 1995], breast, head and neck carcinoma [Okada et al., 1995].

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 [Pei & Weiss, 1996].

MT1-MMP activates progelatinase A [Sato et al., Strongin et al., 1995; Will et al., 1996] and procollagenase-3 [Knauper et al., 1996] 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 ¾ and ¼ fragments [D′Ortho et al., 1997; Ohuci et al., 1997] and it cleaves a number of other ECM proteins, including fibronectin, vitronectin, laminin-1 and dermatan sulfate proteoglycan [D′Ortho et al., 1997; Pei & Weiss, 1996; Ohuci et al., 1997]. The activity of MT1-MMP is poorly inhibited by TIMP-1 but efficiently inhibited by TIMP-2 and TIMP-3 [Will et al., 1996].

CC1043 is a recombinant polypeptide sequence produced as a periplasmic protein in E. coli. The proenzyme consists of MT1-MMP residues corresponding to Ser1-Val501 followed by tone Thr-residue and six His-residues. The calculated Mr of the recombinant soluble proenzyme is 58200 Da.

Useful as an antigen standard in immunoassays. The proenzyme can be activated with trace amounts of MT1-MMP catalytic domain (D′Ortho et al., 1997; Butler et al., 1998).

Provided as a liquid in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2.

Maintain frozen at -70°C in undiluted aliquots. The enzyme may be stored at -20°C for several weeks. Repeated freezing and thawing should be avoided

Appears as a predominant band at 58 kDa in SDS-PAGE

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