Dynamic Interdomain Interactions Contribute to the Inhibition of Matrix Metalloproteinases by Tissue Inhibitors of Metalloproteinases

Abstract

Because of their important function, matrix metalloproteinases (MMPs) are promising drug targets in multiple diseases, including malignancies. The structure of MMPs includes a catalytic domain, a hinge, and a hemopexin domain (PEX), which are followed by a transmembrane and cytoplasmic tail domains or by a glycosylphosphatidylinositol linker in membrane-type MMPs (MT-MMPs). TIMPs-1, -2, -3, and -4 are potent natural regulators of the MMP activity. These are the inhibitory N-terminal and the non-inhibitory C-terminal structural domains in TIMPs. Based on our structural modeling, we hypothesized that steric clashes exist between the non-inhibitory C-terminal domain of TIMPs and the PEX of MMPs. Conversely, a certain mobility of the PEX relative to the catalytic domain is required to avoid these obstacles. Because of its exceedingly poor association constant and, in contrast with TIMP-2, TIMP-1 is inefficient against MT1-MMP. We specifically selected an MT1-MMP⅐TIMP-1 pair to test our hypothesis, because any improvement of the inhibitory potency would be readily recorded. We characterized the domain-swapped MT1-MMP chimeras in which the PEX of MMP-2 (that forms a complex with TIMP-2) and of MMP-9 (that forms a complex with TIMP-1) replaced the original PEX in the MT1-MMP structure. In contrast with the wild-type MT1-MMP, the diverse proteolytic activities of the swapped-PEX chimeras were then inhibited by both TIMP-1 and TIMP-2. Overall, our studies suggest that the structural parameters of both domains of TIMPs have to be taken into account for their re-engineering to harness the therapeutic in vivo potential of the novel TIMP-based MMP antagonists with constrained selectivity.There are 24 individual MMPs 2 in humans. MMPs cleave multiple extracellular matrix components, growth factors, cytokines, and cell signaling adhesion receptors. Aberrant performance of MMPs plays a role in a plethora of diseases, including cancer (1-3). There are 18 soluble and 6 membrane MMPs. The structure of soluble MMPs includes a prodomain (PRO), a catalytic domain (CAT) that contains the active site zinc, a hinge, and a PEX. Additionally, membrane-type MMPs (MT-MMP) contain a transmembrane domain followed by a short cytoplasmic tail (CYTO) (MT1-MMP, MT2-MMP, MT3-MMP, and MT5-MMP) or a glycosylphosphatidylinositol moiety (MT4-MMP and MT6-MMP) that anchors the proteinase to the cell surface (1, 4, 5).MMPs are synthesized as zymogens, which require the proteolytic processing of the N-terminal inhibitory PRO to generate the active enzymes (4). It is accepted that secretory tissue inhibitors of MMPs (TIMPs) play an important role in the regulation of the proteolytic activity of MMPs (6, 7). Four TIMPs (TIMP-1, -2, -3, and -4) are present in humans (8). There is at least a 25% sequence identity among all TIMPs, including 12 conserved Cys residues that form 6 disulfide bridges resulting in 6 loop regions. There are two domains, N-terminal and C-terminal, in TIMPs. An N-terminal domain (NT-TIMP) binds the CAT, carries the MMP-inhibitory activ...

Non-destructive and Selective Imaging of the Functionally Active, Pro-invasive Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) Enzyme in Cancer Cells

“…Effects of TIMPs-To corroborate our results further, we tested if TIMP-1 and TIMP-2 (an inefficient and a very potent inhibitor of MT1-MMP, respectively) affected the binding of MP-3653 to cellular MT1-MMP (35,36,68). For these purposes, MCF7-MT1 cells were co-incubated with TIMP-1 or TIMP-2 (at a 40:1 TIMP-MP-3653 molar ratio) prior to adding MP-3653.…”

Rubiarbonone C inhibits platelet‐derived growth factor‐induced proliferation and migration of vascular smooth muscle cells through the focal adhesion kinase, MAPK and STAT3 Tyr705 signalling pathways

“…We also confirmed that rubiarbonone C inhibited PDGF BB‐induced VSMC migration via down‐regulation of both the expression and activity of MMP2 and 9 (Figure ). Thus, rubiarbonone C may inhibit cell migration by inhibiting external substrate degradation mediated by MMP2 and 9 (Remacle et al, ). The activation of FAK has also been suggested to play an important role in the reorganization of the cytoskeleton for cell movement (Wu et al, ).…”

Promotion of astrocytoma cell invasion by micro RNA–22 targeting of tissue inhibitor of matrix metalloproteinase–2

“…Various proteinases participate in the degradation of ECM, and a family of MMPs plays an important role in matrix degradation around tumor cells. 4,23 A significant correlation between the malignancy of glioma and MMPs has been reported. 17 The activities of MMPs are strictly regulated by TIMPs, 30 which comprise 2 structural domains.…”

“…4 Binding of the C-terminal domain is essential for cell surface activation of MMP-2 by MMP-14, 14,33 and TIMP-2 may play a role in the degradation of the ECM and regulation of glioma invasion. 23 A correlation between miRNA and glioma invasion has been reported, with some miRNAs being involved in the invasion by regulating the degradation of the ECM. Whereas miR-21 promotes glioma invasiveness by targeting MMP inhibitors, 5 miR-132 inhibits glioma cell invasion by targeting MMP16, 31 and miR-23a promotes invasion by modulating MMP-14.…”