1.56 Å structure of mature truncated human fibroblast collagenase

Spurlino, John; Smallwood, Angela; Carlton, Dennis; Banks, Tracey M.; Vavra, Karen J.; Johnson, Jeffrey S.; Cook, Ewell R.; Falvo, Joseph; Wahl, R.; Pulvino, Tricia A.; Wendoloski, John J.; Smith, Douglas L.

doi:10.1002/prot.340190203

Cited by 199 publications

(194 citation statements)

References 23 publications

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“…Three of the Asn double-clash examples are consistent with the possibility of chemical de-amidation of the side-chain: 1CSE Asn158E (subtilisin; Bode et al, 1987), 1HFC Asn206 (collagenase; Spurlino et al, 1994), and 1LUC Asn12 (luciferase; Fisher et al, 1996), which is shown in Figure 8. The tight interactions around Asn12, including two H-bonds to backbone NH groups, de®nitely require an O d for the left-hand branch of the amide.…”

Section: Summary Of Final Assignmentssupporting

confidence: 58%

Asparagine and glutamine: using hydrogen atom contacts in the choice of side-chain amide orientation 1 1Edited by J. Thornton

Word

Lovell

Richardson

et al. 1999

Journal of Molecular Biology

1,339

1,150

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Section: Summary Of Final Assignmentssupporting

confidence: 58%

Asparagine and glutamine: using hydrogen atom contacts in the choice of side-chain amide orientation 1 1Edited by J. Thornton

Word

Lovell

Richardson

et al. 1999

Journal of Molecular Biology

1,339

1,150

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“…These peptides were chosen based on knowledge gained from structure-function studies performed on other MMPs. The structures for the catalytic domains of several members of the MMP family have recently become available including human fibroblast collagenase (MMP-1) (22-24), human stromelysin-1 (MMP-3) (25,26), human matrilysin (MMP-7) (27), and human neutrophil collagenase (MMP-8) (28 -30). These structures revealed that the SЈ 1 subsite is the most well defined pocket in these MMPs and consists of a hydrophobic pocket which varies greatly in its depth.…”

Section: Figmentioning

confidence: 99%

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

Gronski

Martin²,

Kobayashi

et al. 1997

Journal of Biological Chemistry

275

204

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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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“…How collagenases interact with collagen and how the Hpx domain endows these enzymes with collagenolytic activity is not clearly understood. Another enigma of collagenolysis became apparent when the crystal structures of MMP-1Cat (8)(9)(10), MMP-8Cat (11,12), and full-length pig MMP-1 (6) were solved: The catalytic cleft of these enzymes is too narrow to accommodate collagen in its native triple-helical conformation. A number of hypotheses have been proposed to explain how collagenases may destabilize and cleave triple-helical collagen (13)(14)(15)(16), and we have experimentally demonstrated that MMP-1 unwinds triple-helical collagen locally before peptide bond hydrolysis (17,18).…”

mentioning

confidence: 99%

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

Manka

Carafoli

Visse

et al. 2012

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

191

310

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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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1.56 Å structure of mature truncated human fibroblast collagenase

Cited by 199 publications

References 23 publications

Asparagine and glutamine: using hydrogen atom contacts in the choice of side-chain amide orientation 1 1Edited by J. Thornton

Asparagine and glutamine: using hydrogen atom contacts in the choice of side-chain amide orientation 1 1Edited by J. Thornton

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

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

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