The Role of the C-terminal Domain of Human Collagenase-3 (MMP-13) in the Activation of Procollagenase-3, Substrate Specificity, and Tissue Inhibitor of Metalloproteinase Interaction

Abstract

). The kinetics of activation were not affected by removal of the hemopexin-like C-terminal domain. The specific activities of both collagenase-3 and ⌬ 249 -451 collagenase-3 were found to be similar using two quenched fluorescent substrates, but ⌬ 249 -451 collagenase-3 failed to cleave native triple helical collagens (types I and II) into characteristic one-and three-quarter fragments. It was noted, however, that the ␤1,2(I) chains of type I collagen were susceptible to ⌬ 249 -451 collagenase-3, which indicates that the catalytic domain displays telopeptidase activity, thereby generating ␣1,2(I) chains that are slightly shorter than those in native type I collagen. It can be concluded that the C-terminal domain is only essential for the triple helicase activity of collagenase-3. Binding of procollagenase-3 and active collagenase-3 to type I collagen is mediated by the C-terminal domain. Both collagenase-3 and ⌬ 249 -451 collagenase-3 hydrolyzed the large tenascin C isoform, fibronectin, recombinant fibronectin fragments, and type IV, IX, X, and XIV collagens; thus, these events were independent from C-terminal domain interactions. In contrast, the minor cartilage type XI collagen was resistant to cleavage. Kinetic analysis of the mechanism of inhibition of wild-type and ⌬ 249 -451 collagenase-3 by wild-type and mutant tissue inhibitors of metalloproteinase (TIMPs) revealed that the association rates for complex formation were influenced by both Nand C-terminal domain interactions. The C-terminal domain of wild-type collagenase-3 promoted increased association rates with the full-length inhibitors TIMP-1 and TIMP-3 and the hybrid N.TIMP-2/C.TIMP-1 by a factor of up to 33. In contrast, the association rates for complex formation with TIMP-2 and N.TIMP-1/C.TIMP-2 were only marginally affected by C-terminal domain interactions.The matrix metalloproteinases (MMPs) 1 are a family of zincdependent enzymes that have the capacity to degrade most protein components of the extracellular matrix. Their uncontrolled activity contributes to the tissue destruction that is observed during such diverse pathologies as arthritis and cancer. Four main subfamilies of MMPs have been defined according to their substrate specificity, primary structures, and cellular localization: the collagenases, stromelysins, gelatinases, and membrane-type matrix metalloproteinases.Human procollagenase-3 (MMP-13) is a new member of the collagenase subfamily of MMPs, which consists of three members showing an overall sequence homology of 55% (1-4). Human collagenase-3 expression has been demonstrated in breast tumors and in osteoarthritic cartilage (1, 5), indicating that the enzyme plays a role in degradation of collagen during disease progression. Furthermore, its expression in cartilage was strongly induced at both the message and protein levels by interleukin-1␣. Biochemical characterization of human collagenase-3 has shown that it is a powerful collagenolytic enzyme, preferentially cleaving type II collagen, while it is five or six times less ...

Study of matrix metalloproteinases and their inhibitors in breast cancer

“…Matrix metalloproteinase -13 (collagenase-3) has been found to have an exceptionally wide substrate specificity when compared with other MMPs (Freije et al, 1994;Knauper et al, 1997). Moreover, it is thought to play a central role in the MMP activation cascade, both activating and being activated by several other MMPs (MMP-14, 2, or 3).…”

Inhibition of collagenase-3 (MMP-13) expression in transformed human keratinocytes by interferon-γ is associated with activation of extracellular signal-regulated kinase-1,2 and STAT1

“…In addition, TNF-a and TGF-b-elicited stimulation of A-5 cell invasion through Matrigel was entirely abrogated by IFN-g most likely due to inhibition of MMP-9 production. However, as MMP-13 also degrades type IV collagen (KnaÈ uper et al, 1997), it is possible that reduction in MMP-13 production also impairs the ability of transformed keratinocytes to invade through basement membrane. Together these observations clearly show, that IFN-g plays an important role in limiting the invasion capacity of transformed keratinocytes.…”

Heterogeneous requirement of IκB kinase 2 for inflammatory cytokine and matrix metalloproteinase production in rheumatoid arthritis: Implications for therapy

“…We subsequently investigated the role of IKK-2 in the production of MMPs in the RA synovial membrane. These enzymes play a role in the destructive process of RA by breaking down human cartilage and bone (54)(55)(56). We previously showed that MMP-1 and MMP-3 expression in ex vivo RA synovial membranes is NF-B dependent because AdI B␣ down-regulates their expression (17).…”