Flexibility and Variability of TIMP Binding: X-ray Structure of the Complex Between Collagenase-3/MMP-13 and TIMP-2

Maskos, K.; Lang, R.; Tschesche, Harald; Bode, Wolfram

doi:10.1016/j.jmb.2006.11.072

Cited by 50 publications

(57 citation statements)

References 54 publications

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“…Crystal structures of multiple TIMP⅐MMP inhibitory complexes have been determined. They demonstrate similar inhibitory mechanisms, involving a region surrounding the conserved Cys 1 -Cys 70 disulfide bond of TIMPs and especially the N-terminal Cys 1 -Pro 6 and Glu 67 -Cys 70 segments (numbering is given according to TIMP-1) that approach the MMP active site (PDB 1UEA, 1BUV, 1GXD, 1BR9, and 2E2D) (12)(13)(14)(15)(16)(17)(18). The Cys 1 coordinates, via bidentate interactions, the catalytic zinc of the MMP active site.…”

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confidence: 98%

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

Remacle

Shiryaev

Radichev

et al. 2011

Journal of Biological Chemistry

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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...

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mentioning

confidence: 98%

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

Remacle

Shiryaev

Radichev

et al. 2011

Journal of Biological Chemistry

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“…Each Srt variant was characterized with respect to secondary structure content by CD spectroscopy. The far UV CD spectra of STX and its Val 16 , Ala 4 , and Ser 4 variants (Fig. 5) resemble that of full-length Srt b (22,45).…”

Section: Resultsmentioning

confidence: 81%

“…1 (14). Similar contacts were observed subsequently in the structure of complexes of TIMP-2 with MMP-14 (15) and MMP-13 (16) and the complex of N-TIMP-1 with MMP-1 (17). To develop their potential as therapeutic agents while minimizing deleterious side effects, N-TIMPs have been successfully engineered by mutation of their MMP interaction sites to enhance their inhibitory selectivity for different metalloproteinases (18 -20).…”

mentioning

confidence: 57%

“…2). The C-terminal region is analogous to residues 65-69 of TIMPs (numbering refers to human TIMP-1), which forms part of the TIMP/MMP contact surface (11,(15)(16)(17)46) but is oriented in the opposite direction. Using sequence information from 13 different TIMPs, a limited combinatorial peptide synthesis scheme was devised.…”

Section: Resultsmentioning

confidence: 99%

“…It contained the C-terminal sequence Met-Ser-Glu-Met-Ser, which is not identical with the "equivalent" sequence in any known TIMP. Interestingly, Met 16 and Ser 19 correspond to nonmammalian TIMPs (Drosophila and Caenorhabditis elegans), which were deliberately underrepresented in our original combinatorial library. This peptide was synthesized, folded, purified, and screened for metalloproteinase inhibitory activity; it was found to be a 10 -30-fold better inhibitor than STX-S4 (Table 2 and Fig.…”

Section: Resultsmentioning

confidence: 99%

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Engineered Sarafotoxins as Tissue Inhibitor of Metalloproteinases-like Matrix Metalloproteinase Inhibitors

Lauer‐Fields

Čudić

Wei

et al. 2007

Journal of Biological Chemistry

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The sarafotoxins and endothelins are ϳ25-residue peptides that spontaneously fold into a defined tertiary structure with specific pairing of four cysteines into two disulfide bonds. Their structures show an interesting topological similarity to the core of the metalloproteinase interaction sites of the tissue inhibitors of metalloproteinases. Previous work indicates that sarafotoxins and endothelins can be engineered to eliminate or greatly reduce their vasopressive action and that their structural framework can withstand multiple sequence changes. When sarafotoxin 6b, which possesses modest matrix metalloproteinase inhibitory activity, was C-terminally truncated to remove its toxic vasopressive activity, the metalloproteinase inhibitory activity was essentially abolished. However, further changes, based on the sequences of peptides selected from libraries of sarafotoxin variants or suggested by analogy with tissue inhibitors of metalloproteinases, progressively enhanced the matrix metalloproteinase inhibitory activity. Peptide variants with multiple substitutions folded correctly and formed native disulfide bonds. Improvements in matrix metalloproteinase affinity have generated a peptide with micromolar K i values for matrix metalloproteinase-1 and -9 that are selective inhibitors of different metalloproteinases. Characterization of its solution structure indicates a close similarity to sarafotoxin but with a more extended C-terminal helix. The effects of N-acetylation and other changes, as well as docking studies, support the hypothesis that the engineered sarafotoxins bind to matrix metalloproteinases in a manner analogous to the tissue inhibitors of metalloproteinases. The endothelins (ET)3 are a family of 21-25-residue peptides that fall into two major categories, endothelins and sarafotoxins (Srt). The names are derived from their biological sources, endothelial cells, and the venoms of snakes of the genus Atractaspis (in Hebrew, Saraf Ein Gedi). Three ETs are found in humans, and an array of Srts are present in the venoms of different Atractaspis sp. ET family complexity arises from a variety of mechanisms; expression from multiple genes, proteolytic processing, and possibly RNA editing (1-4). NMR and crystallographic studies reveal that these peptides contain a bicyclic two-disulfide (Cys 1 -Cys 15 , Cys 3 -Cys 11 ) framework with a specific tertiary structure. The N-terminal residues form a ␤-strand followed by a ␤-turn at residue 5 (5); residues 9 -16 are mainly ␣-helical, whereas the C-terminal residues are flexible in solution (6). However, the crystal structure and protease susceptibility studies suggest that these C-terminal residues may associate with the bicyclic core as an ␣-helix (7, 8).ETs have many physiological effects including stimulation of cell proliferation, promotion of the expression of cell adhesion molecules and extracellular matrix components, activation of the inflammatory cascade, and the well characterized contraction of smooth muscle tissue (9). In humans, these biological acti...

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Converting a broad matrix metalloproteinase family inhibitor into a specific inhibitor of MMP‐9 and MMP‐14

et al. 2018

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MMP-14 and MMP-9 are two well-established cancer targets for which no specific clinically relevant inhibitor is available. Using a powerful combination of computational design and yeast surface display technology, we engineered such an inhibitor starting from a nonspecific MMP inhibitor, N-TIMP2. The engineered purified N-TIMP2 variants showed enhanced specificity toward MMP-14 and MMP-9 relative to a panel of off-target MMPs. MMP-specific N-TIMP2 sequence signatures were obtained that could be understood from the structural perspective of MMP/N-TIMP2 interactions. Our MMP-9 inhibitor exhibited 1000-fold preference for MMP-9 vs. MMP-14, which is likely to translate into significant differences under physiological conditions. Our results provide new insights regarding evolution of promiscuous proteins and optimization strategies for design of inhibitors with single-target specificities.

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Flexibility and Variability of TIMP Binding: X-ray Structure of the Complex Between Collagenase-3/MMP-13 and TIMP-2

Cited by 50 publications

References 54 publications

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

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

Engineered Sarafotoxins as Tissue Inhibitor of Metalloproteinases-like Matrix Metalloproteinase Inhibitors

Converting a broad matrix metalloproteinase family inhibitor into a specific inhibitor of MMP‐9 and MMP‐14

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