Stromelysin‐1: Three‐dimensional structure of the inhibited catalytic domain and of the C‐truncated proenzyme

Becker, Joseph W.; Marcy, Alice I.; Rokosz, Laura L.; Axel, Melinda G.; Burbaum, Jonathan J.; Fitzgerald, Paul J.; Cameron, Patricia M.; Esser, Craig K.; Hagmann, William K.; Hermes, Jeffrey D.; Springer, James P.

doi:10.1002/pro.5560041002

Cited by 291 publications

(273 citation statements)

References 61 publications

(45 reference statements)

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“…S7). Some of these hydrogen bonds are identical to those formed with a cysteine-switch sequence (26)(27)(28)(29) in proMMPs and with a predicted peptide substrate (30).…”

Section: Resultsmentioning

confidence: 68%

Structural insights into triple-helical collagen cleavage by matrix metalloproteinase 1

Manka

Carafoli

Visse

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

187

306

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Collagenases of the matrix metalloproteinase (MMP) family play major roles in morphogenesis, tissue repair, and human diseases, but how they recognize and cleave the collagen triple helix is not fully understood. Here, we report temperature-dependent binding of a catalytically inactive MMP-1 mutant (E200A) to collagen through the cooperative action of its catalytic and hemopexin domains. Contact between the two molecules was mapped by screening the Collagen Toolkit peptide library and by hydrogen/deuterium exchange. The crystal structure of MMP-1(E200A) bound to a triplehelical collagen peptide revealed extensive interactions of the 115-Å-long triple helix with both MMP-1 domains. An exosite in the hemopexin domain, which binds the leucine 10 residues C-terminal to the scissile bond, is critical for collagenolysis and represents a unique target for inhibitor development. The scissile bond is not correctly positioned for hydrolysis in the crystallized complex. A productive binding mode is readily modeled, without altering the MMP-1 structure or the exosite interactions, by axial rotation of the collagen homotrimer. Interdomain flexing of the enzyme and a localized excursion of the collagen chain closest to the active site, facilitated by thermal loosening of the substrate, may lead to the first transition state of collagenolysis. extracellular matrix | X-ray crystallography | protease T he interstitial collagens I, II, and III are the major structural proteins in connective tissues such as skin, bone, cartilage, tendon, and blood vessels (1). They consist of three α chains with repeating Gly-X-Y triplets (X and Y are often proline and hydroxyproline, respectively) that intertwine each other to form a triple helix of ∼300 nm in length (2). Interstitial collagens are resistant to most proteolytic enzymes, but vertebrate collagenases cleave them at a single site approximately three-quarters of the way from the N terminus of the triple helix, thus initiating collagenolysis (3). Owing to this unique activity, collagenases play important roles in embryo development, morphogenesis, tissue remodeling, wound healing, and human diseases, such as arthritis, cancer, and atherosclerosis (4, 5).Matrix metalloproteinase 1 (MMP-1) is a typical vertebrate collagenase (3). It consists of an N-terminal catalytic (Cat) domain containing an active-site zinc ion and a C-terminal hemopexin (Hpx) domain comprised of a four-bladed β-propeller, which are connected by a linker region (6, 7). Although the Cat domain can cleave a number of noncollagenous proteins including heat-denatured collagen (gelatin), its activity on native triplehelical collagen is negligible. The combination of the Cat and Hpx domains is required for MMP-1 to be able to degrade native collagen, and the same is true for all other collagenolytic MMPs, namely MMP-2, MMP-8, MMP-13, and MMP-14 (3). How collagenases interact with collagen and how the Hpx domain endows these enzymes with collagenolytic activity is not clearly understood. Another enigma of collagenolysis bec...

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“…S7). Some of these hydrogen bonds are identical to those formed with a cysteine-switch sequence (26)(27)(28)(29) in proMMPs and with a predicted peptide substrate (30).…”

Section: Resultsmentioning

confidence: 68%

Structural insights into triple-helical collagen cleavage by matrix metalloproteinase 1

Manka

Carafoli

Visse

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

187

306

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“…Even so, the estimates of its secondary structure as determined by C.D. (high fl-sheet and negligible a-helix content) are only partly in accord with the recently published crystal structures of porcine fibroblast collagenase [23] and a deletion mutant of human prostromelysin 1 [24]. The C-terminal domain of collagenase is predominantly fl-sheet but both structures have regions of a-helix in their catalytic domains that are made up of sequences similar to those found in human gelatinase A.…”

Section: Discussionmentioning

confidence: 89%

“…Our results also suggest that the propeptide of progelatinase A contains some fl-sheet. This would make it markedly different to the propeptide of prostromelysin-1, which has 3 short a-helices but no fl-sheet [24].…”

Section: Discussionmentioning

confidence: 96%

An analysis of the conformational changes that accompany the activation and inhibition of gelatinase A

Crabbe¹,

Kelly

Price

1996

FEBS Letters

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The latent precursors of the matrix metalloproteinases (MMPs) are converted by (4-aminophenylmercuric)acetate to active forms that lose their propeptide as a result of autolysis. C.D. and an active site mutant of progelatinase A (MMP2) were used to demonstrate that, although propeptide removal is accompanied by a decrease in the enzyme's /]-sheet content, the initial activation is achieved with only minor modifications to the conformation. Mixing activated gelatinase A with the natural inhibitor, TIMP-1, resulted in conformational changes that were absent when a synthetic inhibitor was used. The relevance of these results to MMP activation and inhibition is discussed.

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“…The second site is within the active site loop at the Phe 352 -Leu 353 bond. This is the same site at which interstitial collagenase and the 92-kDa gelatinase cleave ␣ 1 -AT (20,21). Surprisingly, however, MME, the murine orthologue to HME, cleaves at the Pro 357 -Met 358 bond instead.…”

Section: Hme and Elastolysis 12192mentioning

confidence: 95%

Hydrolysis of a Broad Spectrum of Extracellular Matrix Proteins by Human Macrophage Elastase

Gronski

Martin²,

Kobayashi

et al. 1997

Journal of Biological Chemistry

275

203

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Macrophage elastase (ME) was originally named when metal-dependent elastolytic activity was detected in conditioned media of murine macrophages. Subsequent cDNA cloning of the mouse and human enzyme demonstrated that ME is a distinct member of the matrix metalloproteinase family. To date, the catalytic parameters that describe the hydrolysis of elastin by ME have not been quantified and its activity against other matrix proteins have not been described. In this report, we have examined the action of purified recombinant human ME (rHME), produced in Escherichia coli, on elastin and other extracellular matrix proteins. On a molar basis, rHME is approximately 30% as active as human leukocyte elastase in solubilizing elastin. rHME also efficiently degrades ␣ 1 -antitrypsin (␣ 1 -AT), the primary physiological inhibitor of human leukocyte elastase. In addition, rHME efficiently degrades fibronectin, laminin, entactin, type IV collagen, chondroitan sulfate, and heparan sulfate. These results suggest that HME may be required for macrophages to penetrate basement membranes and remodel injured tissue during inflammation. Moreover, abnormal expression of HME may contribute to destructive processes such as pulmonary emphysema and vascular aneurysm formation. To further understand the specificity of HME, the initial cleavage sites in ␣ 1 -AT have been determined. In addition, the hydrolysis of a series of synthetic peptides with different P 1 residues has been determined. rHME can accept large and small amino acids at the P 1 site, but has a preference for leucine.Macrophage elastase (MMP-12) shares many properties with other members of the matrix metalloproteinase (MMP) 1 gene family, yet it is unique in several ways. Like other MMPs, metalloelastase requires zinc for catalytic activity, is inhibited by the tissue inhibitors of metalloproteinases (TIMPs), and has common structural domains with other MMPs (1, 2). Human macrophage elastase (HME) is most closely related to collagenase-1 (MMP-1) and stromelysin-1 (MMP-3), being 49% identical to each at the amino acid level (3). Moreover, the gene for macrophage elastase, composed of a common 10-exon, 9-intron structure, is on human chromosome 11q22.2/22.3 with at least six other MMPs (4). Despite these similarities, HME possesses certain distinct biological and biochemical properties. Expression appears to be largely restricted to tissue macrophages (4). Upon activation, it not only cleaves its 8-kDa N-terminal domain, but also has a unique propensity to autolytically release its 23-kDa C-terminal domain resulting in a mature active 22-kDa proteinase (3)(4)(5).Macrophage elastase shares its elastolytic activity (6, 7) with only a few MMPs, including the gelatinases (MMP-2 and MMP-9) and, to a lesser extent, matrilysin (8, 9). However, despite this characteristic activity, the relative capacity of metalloelastase (human or mouse) to degrade elastin has never been quantified. In addition, the catalytic capacity of metalloelastase against other extracellular matrix components has...

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Stromelysin‐1: Three‐dimensional structure of the inhibited catalytic domain and of the C‐truncated proenzyme

Cited by 291 publications

References 61 publications

Structural insights into triple-helical collagen cleavage by matrix metalloproteinase 1

Structural insights into triple-helical collagen cleavage by matrix metalloproteinase 1

An analysis of the conformational changes that accompany the activation and inhibition of gelatinase A

Hydrolysis of a Broad Spectrum of Extracellular Matrix Proteins by Human Macrophage Elastase

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