Crystal structure of human macrophage elastase (MMP-12) in complex with a hydroxamic acid inhibitor 1 1Edited by I. Wilson

Nar, Herbert; Werle, Karlheinz; Bauer, Margit; Dollinger, Horst; Jung, Birgit

doi:10.1006/jmbi.2001.4953

Cited by 79 publications

(94 citation statements)

References 30 publications

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“…However, even by using a molecular model describing the potential binding mode of 1 within the hMMP-12 active site, it would be hard to predict the results reported in this study. The main reason for this is the fact that Lys 241 is located on a loop segment of hMMP-12, which, based on both x-ray and NMR studies of free hMMP-12 or in complex with synthetic inhibitors, displays high flexibility with the lysine side chain exhibiting high mobility (33)(34)(35). Superimposition of two hMMP-12 structure models, taken from an ensemble of twenty NMR-derived free hMMP-12 structures (34), provides some clues about the conformational space sampled by the Lys 241 side chain (Fig.…”

Section: Discussionmentioning

confidence: 99%

Molecular Determinants of Matrix Metalloproteinase-12 Covalent Modification by a Photoaffinity Probe

Dabert-Gay

Czarny

Devel

et al. 2008

Journal of Biological Chemistry

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Mass spectroscopy, microsequencing, and site-directed mutagenesis studies have been performed to identify in human matrix metalloelastase (hMMP-12) residues covalently modified by a photoaffinity probe, previously shown to be able to covalently label specifically the active site of matrix metalloproteinases (MMPs). Results obtained led us to conclude that photoactivation of this probe in complex with hMMP-12 affects a single residue in human MMP-12, Lys 241 , through covalent modification of its side chain ⑀ NH 2 group. Because x-ray and NMR studies of hMMP-12 indicate that Lys 241 side chain is highly flexible, our data reveal the existence of particular Lys 241 side-chain conformation in which the ⑀ NH 2 group points toward the photolabile group of the probe, an event explaining the high levels of cross-linking yield between hMMP-12 and the probe. Lys 241 is not conserved in MMPs, thus differences in cross-linking yields observed with this probe between MMP members may be linked to the residue variability observed at position 241 in this family.

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Section: Discussionmentioning

confidence: 99%

Molecular Determinants of Matrix Metalloproteinase-12 Covalent Modification by a Photoaffinity Probe

Dabert-Gay

Czarny

Devel

et al. 2008

Journal of Biological Chemistry

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“…This mutation enables NMR studies of MMP-12 without turnover of substrate, without inhibitor, and without autolysis (36,58). The lack of structural perturbation by the mutation beyond the side chain itself is evident from comparison of crystal structures (9,30) and from the minimal and localized chemical shift perturbations of NMR spectra (36). Recombinant E219A-inactivated catalytic domain was prepared as described previously (36,58,59 NMR Spectroscopy-NMR spectra and titrations were acquired at 26°C on a Varian Inova 600-MHz spectrometer equipped with a high sensitivity 5-mm cryogenic HCN triple resonance probe with an actively shielded z-gradient coil.…”

Section: Methodsmentioning

confidence: 99%

MMP-12 Catalytic Domain Recognizes Triple Helical Peptide Models of Collagen V with Exosites and High Activity

Bhaskaran

Palmier

Lauer‐Fields

et al. 2008

Journal of Biological Chemistry

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Collagens are principal constituents of connective tissues. They are hydrolyzed during development, wound repair, and remodeling of the extracellular matrix. Dysregulated collagenolysis is active in inflammatory diseases such as atherosclerosis, cancer, rheumatoid arthritis, periodontitis, and liver and kidney pathologies. Matrix metalloproteinases (MMPs) 3 play key roles in these processes (1-3). Mutagenesis has established the importance of the V-B loop (joining ␤-strand V and helix B) in collagenolysis by MMP-1 (4). Residues at the C-terminal end of this loop in MMP-8 were also implicated in collagenolysis (5) and triple helical peptidase activity (6).Binding of an inactivated MMP-1 to an intact collagen fibril likely unwinds the triple helix at the cleavage site to provide access for an active MMP-1 catalytic domain to hydrolyze individual chains (7). The direction of collagen triple helices at the active site is unknown, but the orientation can be hypothesized to be the same as in linear peptide substrates that run antiparallel to ␤-strand IV (8, 9). Single chain peptides from the ␣1 chain of type I/III collagens were soaked into crystals of MMP-12 and MMP-8 catalytic domains, and snapshots of bound hydrolysis products were obtained (8).However, neither crystallography nor NMR data representative of interactions with collagen have been reported because of technical limitations. More generally, structural studies of complexes of enzymes with their substrates have been rare presumably because of catalytic turnover and affinities lower than those for transition state analogues. Several biologically active, self-assembling triple helical peptide (THP) mimics of collagens have been developed. These THPs have facilitated numerous biochemical and cell biological studies otherwise infeasible with insoluble, intact collagen fibrils. THPs are successful in reproducing the cleavage sites of MMPs in collagens I, II, III, V, and XI and in reproducing rate dependences on intact triple helical structure (10, 11). Fluorogenic labeling of THPs has facilitated quantitative comparisons of the triple helical peptidase activities of a variety of MMPs in solution and in cell culture assay (6,11,12). The affinities of MMP catalytic domains for triple helices range from 1 to 80 M (6, 11-13). Collagenolysis involves several MMP functions that include initial nonspecific binding and orientation of collagen fibrils (14, 15), manipulation of collagen fibrils with the C-terminal hemopexin domain (16) and catalytic domain (7), unwinding of triple helices within fibrils, and ensuing hydrolysis of single chains from the melted triple helix (7). THPs are not suitable for modeling the initial binding, orientation, and manipulation of full-length collagen assembled into large, insoluble fibrils (6, 11). They are too small to participate in these higher order events (7). Nonetheless THPs have proven themselves to be outstanding substrates for detailed characterization of the triple helical peptidase activities of MMPs and other metalloprotein...

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“…MMI270 is a hydroxamate inhibitor that inhibits MMP-2, -3, -9, -14, and -12 at nanomolar concentrations. 37,38 This compound binds the S1Ј pocket of MMP enzymes, 39 affording it broader substrate specificity than BAY-129566. This activity underlies the clinical application of the compound, which is primarily aimed at lung fibrosis where macrophage-derived MMP-12 has been shown to underlie fibrogenesis.…”

Section: Mmp-12 Expression Is Markedly Induced In Glomeruli From Alpomentioning

confidence: 99%

Role for Macrophage Metalloelastase in Glomerular Basement Membrane Damage Associated with Alport Syndrome

Rao

Meehan

Delimont

et al. 2006

The American Journal of Pathology

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Crystal structure of human macrophage elastase (MMP-12) in complex with a hydroxamic acid inhibitor 1 1Edited by I. Wilson

Cited by 79 publications

References 30 publications

Molecular Determinants of Matrix Metalloproteinase-12 Covalent Modification by a Photoaffinity Probe

Molecular Determinants of Matrix Metalloproteinase-12 Covalent Modification by a Photoaffinity Probe

MMP-12 Catalytic Domain Recognizes Triple Helical Peptide Models of Collagen V with Exosites and High Activity

Role for Macrophage Metalloelastase in Glomerular Basement Membrane Damage Associated with Alport Syndrome

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