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F X Gomisruth - One of the best experts on this subject based on the ideXlab platform.

  • the metzincins topological and sequential relations between the astacins Adamalysins serralysins and matrixins collagenases define a superfamily of zinc peptidases
    Protein Science, 2008
    Co-Authors: Walter Stocker, F X Gomisruth, Ulrich Baumann, Frank Grams, Peter Reinemer, David B Mckay, Wolfram Bode
    Abstract:

    The three-dimensional structures of the zinc endopeptidases human neutrophil collagenase, Adamalysin II from rattle snake venom, alkaline proteinase from Pseudomonas aeruginosa, and astacin from crayfish are topologically similar, with respect to a five-stranded beta-sheet and three alpha-helices arranged in typical sequential order. The four proteins exhibit the characteristic consensus motif HEXXHXXGXXH, whose three histidine residues are involved in binding of the catalytically essential zinc ion. Moreover, they all share a conserved methionine residue beneath the active site metal as part of a superimposable "Met-turn." This structural relationship is supported by a sequence alignment performed on the basis of topological equivalence showing faint but distinct sequential similarity. The alkaline proteinase is about equally distant (26% sequence identity) to both human neutrophil collagenase and astacin and a little further away from Adamalysin II (17% identity). The pairs astacin/Adamalysin II, astacin/human neutrophil collagenase, and Adamalysin II/human neutrophil collagenase exhibit sequence identities of 16%, 14%, and 13%, respectively. Therefore, the corresponding four distinct families of zinc peptidases, the astacins, the matrix metalloproteinases (matrixins, collagenases), the Adamalysins/reprolysins (snake venom proteinases/reproductive tract proteins), and the serralysins (large bacterial proteases from Serratia, Erwinia, and Pseudomonas) appear to have originated by divergent evolution from a common ancestor and form a superfamily of proteolytic enzymes for which the designation "metzincins" has been proposed. There is also a faint but significant structural relationship of the metzincins to the thermolysin-like enzymes, which share the truncated zinc-binding motif HEXXH and, moreover, similar topologies in their N-terminal domains.

  • structures of Adamalysin ii with peptidic inhibitors implications for the design of tumor necrosis factor alpha convertase inhibitors
    Protein Science, 1998
    Co-Authors: F X Gomisruth, Lawrence F. Kress, Edgar F Meyer, V Politi
    Abstract:

    Croralus adamanteus snake venom Adamalysin I1 is the structural prototype of the Adamalysin or ADAM family comprising proteolytic domains of snake venom metatloproteinases, multimodular mammalian reproductive tract proteins, and tumor necrosis factor a convertase, TACE, involved in the release of the inflammatory cytokine, TNFa. The structure of Adamalysin I1 in noncovalent complex with two small-molecule right-hand side peptidomimetic inhibitors (Pol 647 and Pol 656) has been solved using X-ray diffraction data up to 2.6 and 2.8 8, resolution. The inhibitors bind to the S’-side of the proteinase, inserting between two protein segments, establishing a mixed parallel-antiparallel three-stranded P-shest and coordinate the central zinc ion in a bidentate manner via their two C-terminal oxygen atoms. The proteinase-inhibitor complexes are described in detail and are compared with other known structures. An Adamalysinbased model of the active site of TACE reveals that these small molecules would probably fit into the active site cleft of this latter metalloproteinase, providing a starting model for the rational design of TACE inhibitors.

  • 2 a x ray structure of Adamalysin ii complexed with a peptide phosphonate inhibitor adopting a retro binding mode
    FEBS Letters, 1997
    Co-Authors: Maurizio Cirilli, Lawrence F. Kress, F X Gomisruth, Carlo Gallina, E Gavuzzo, Cesare Giordano, Barbara Gorini, F Mazza, Paglialunga M Paradisi, Giorgio Pochetti
    Abstract:

    The search of reprolysin inhibitors offers the possibility of intervention against both matrixins and ADAMs. Here we report the crystal structure of the complex between Adamalysin II, a member of the reprolysin family, and a phosphonate inhibitor modeled on an endogenous venom tripeptide. The inhibitor occupies the primed region of the cleavage site adopting a retro-binding mode. The phosphonate group ligates the zinc ion in an asymmetric bidentate mode and the adjacent Trp indole system partly fills the primary specificity subsite S1′. An Adamalysin-based model of tumor necrosis factor-α-converting enzyme (TACE) reveals a smaller S1′ pocket for this enzyme.

  • the crystal structure of Adamalysin ii a zinc endopeptidase from the snake venom of the eastern diamondback rattlesnake crotalus adamanteus
    Brazilian Journal of Medical and Biological Research, 1994
    Co-Authors: Wolfram Bode, Lawrence F. Kress, Edgar F Meyer, F X Gomisruth
    Abstract:

    : 1. Adamalysin II, alias proteinase II, a 24-kDa zinc-endopeptidase from the snake venom of Crotalus adamanteus, is a member of a large family of metalloproteinases isolated as small proteinases or proteolytic domains of mosaic hemorrhagic proteins from various snake venoms. Homologous domains have been recently detected in multimodular mammalian reproductive tract proteins and in mammalian gene products, somatic rearrangements of which seem to be linked to primary breast cancers. 2. The 2.0 A X-ray crystal structure of Adamalysin II reveals an ellipsoidal molecule with a shallow active-site cleft separating a relatively irregularly folded sub-domain from the main molecular body composed of a 5-stranded beta-sheet and four alpha-helices. Opposite to this active-site cleft is an integrated calcium ion liganded by carbonyl and strongly conserved carboxylate/carboxamide residues. The folding of the peptide fragment containing the zinc-binding motif HExxHxxGxxH bears only a distant resemblance to thermolysin; it is identical to that found in astacin, in collagenases, and in serralysins, with the three histidines (His142, His146, His152) and a water molecule (linked to the glutamic acid Glu143) likewise constituting the zinc ligand; similar to collagenases, but in contrast to astacin, Adamalysin II lacks a fifth (tyrosine) zinc ligand, leaving its zinc-ion tetrahedrally coordinated. Furthermore, Adamalysin II shares an identical active-site basement formed by a common Met-turn. 3. Due to their virtually identical active-site environment and similar folding topology, the snake venom metalloproteinases (hitherto called Adamalysins) and the three other proteinases might be grouped into a common superfamily called metzincins with distinct differences from the thermolysin family.

  • refined 2 0 a x ray crystal structure of the snake venom zinc endopeptidase Adamalysin ii primary and tertiary structure determination refinement molecular structure and comparison with astacin collagenase and thermolysin
    Journal of Molecular Biology, 1994
    Co-Authors: F X Gomisruth, L F Kress, Josef Kellermann, Irmgard Mayr, Robert Huber, Wolfram Bode
    Abstract:

    Abstract Adamalysin II, alias proteinase II, a 24 kDa zinc-endopeptidase isolated from the snake venom of the Eastern diamondback rattlesnake Crotalus adamanteus , is a prototype, of the proteolytic domain of snake venom metalloproteinases and of domains found in mammalian reproductive tract proteins. Its 2·0 A crystal and molecular structure was solved by multiple isomorphous replacement using six heavy-atom derivatives, and was refined to a crystallographic R -value of 0·172. 201 of the 203 amino acid residues of Adamalysin II are defined by electron density; only the first two residues are disordered and crystallographically undefined in the crystal structure. Three-quarters of these crystallographic amino acid residue assignments were confirmed by chemical sequencing. In addition, the active-site zinc-ion, a heptaco-ordinated calcium ion, a fixed sulphate anion and 173 solvent molecules were localized in the structure. Adamalysin II is an ellipsoidal molecule with a relatively flat active-site cleft separating the "upper" main body from a small "lower" subdomain. The regularly folded N-terminal upper domain consists essentially of a central, highly twisted five-stranded β-pleated sheet flanked by a long and a short surface located helix on its convex side, and by two long helices, one of which represents the central "active site helix", on its concave side. The lower subdomain, comprising the last 50 residues, is organized in multiple turns, with the chain ending in a long C-terminal helix and an extended segment clamped to the upper domain via a disulphide bridge. The catalytic zinc-ion, located at the bottom of the active-site cleft, is almost tetrahedrally co-ordinated by His142, His146 and His152, and a water molecule anchored to an intermediate glutamic acid residue (Glu143), with the three imidazole Ne nitrogen atoms 2·1 A and the solvent oxygen atom 2·4 A away from the zinc ion. His142, Glu143 and His146 are part of the long active-site helix, which extends up to Gly149, where it turns sharply away towards His152. The importance of these residues for structure and activity of Adamalysin II explains their occurrence in the HEXXHXXGXXH consensus sequence. Asp153, which is strictly conserved in these snake venom and reproductive tract metalloproteinases, is buried in the subdomain and seems to stabilize the hydrophobic active-site basement. Some residues behind, the Adamalysin peptide chain folds into a characteristic 1,4-turn (the "Met-turn") containing the conserved Met166, which forms a hydrophobic basement for the three zinc-binding imidazoles. Adamalysin II shares a similar overall topology with astacin, with the collagenase catalytic domains and the P. aeruginosa alkaline proteinase, and exhibits a virtually identical zinc environment with these other "metzincins". The catalytic domains of the snake venom metalloproteinases and the corresponding domains of some reproductive tract proteins can be aligned with only a very few single residue insertions and deletions and with sequence identities mostly above 50%. All substitutions, including some novel cysteine residues engaged in additional disulphide bridges, are in agreement with the Adamalysin 11 structure, which can therefore be considered as a prototype of these "adarnalysins".

Linda Troeberg - One of the best experts on this subject based on the ideXlab platform.

  • dissecting the interaction between tissue inhibitor of metalloproteinases 3 timp 3 and low density lipoprotein receptor related protein 1 lrp 1 development of a trap to increase levels of timp 3 in the tissue
    Matrix Biology, 2017
    Co-Authors: Simone D Scilabra, Kazuhiro Yamamoto, Martina Pigoni, Kazuma Sakamoto, Stephan A Muller, Alkmini A Papadopoulou, Stefan F Lichtenthaler, Linda Troeberg
    Abstract:

    Tissue inhibitor of metalloproteinases 3 (TIMP-3) is a key regulator of extracellular matrix turnover for its ability to inhibit matrix metalloproteinases (MMPs), Adamalysin-like metalloproteinases (ADAMs) and ADAMs with thrombospondin motifs (ADAMTSs). TIMP-3 is a secreted protein whose extracellular levels are regulated by endocytosis via the low-density-lipoprotein receptor-related protein-1 (LRP-1). In this study we developed a molecule able to "trap" TIMP-3 extracellularly, thereby increasing its tissue bioavailability. LRP-1 contains four ligand-binding clusters. In order to investigate the TIMP-3 binding site on LRP-1, we generated soluble minireceptors (sLRPs) containing the four distinct binding clusters or part of each cluster. We used an array of biochemical methods to investigate the binding of TIMP-3 to different sLRPs. We found that TIMP-3 binds to the ligand-binding cluster II of the receptor with the highest affinity and a soluble minireceptor containing the N-terminal half of cluster II specifically blocked TIMP-3 internalization, without affecting the turnover of metalloproteinases. Mass spectrometry-based secretome analysis showed that this minireceptor, named T3TRAP, selectively increased TIMP-3 levels in the extracellular space and inhibited constitutive shedding of a number of cell surface proteins. In conclusion, T3TRAP represents a biological tool that can be used to modulate TIMP-3 levels in the tissue and could be potentially developed as a therapy for diseases characterized by a deficit of TIMP-3, including arthritis.

  • ADAMTS and ADAM metalloproteinases in osteoarthritis - looking beyond the 'usual suspects'.
    Osteoarthritis and Cartilage, 2017
    Co-Authors: C Y Yang, Anastasios Chanalaris, Linda Troeberg
    Abstract:

    Summary Introduction Matrix metalloproteinases (MMPs) and ‘aggrecanase' a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs) are well established to play key roles in osteoarthritis (OA) through degradation of extracellular matrix (ECM) type II collagen and aggrecan, and are thus potential targets for development of OA therapies. Objective This paper aims to provide a comprehensive review of the expression and potential roles of other, lesser-known ADAMTSs and related Adamalysins (or a disintegrin and metalloproteinases (ADAMs)) in cartilage, with a view to identifying potentially protective or homeostatic metalloproteinases in the joint and informing consequent selective inhibitor design. Design A comprehensive literature search was performed using PubMed terms ‘osteoarthritis' and ‘ADAMTS' or ‘ADAM'. Results Several ADAMTSs and ADAMs were identified as having reportedly increased expression in OA. These include enzymes likely to play roles in cartilage matrix anabolism (e.g., the procollagen N-proteinases ADAMTS-2, ADAMTS-3 and ADAMTS-14), chondrocyte differentiation and proliferation (e.g., ADAM9, ADAM10, ADAM12), as well as enzymes contributing to cartilage catabolism (e.g., Cartilage oligomeric protein (COMP)-degrading ADAMTS-7 and ADAMTS-12). Conclusions In addition to the well-characterised MMPs, ADAMTS-4 and ADAMTS-5, many other ADAMTSs and ADAMs are expressed in cartilage and several show significantly altered expression in OA. Studies aimed at elucidating the pathophysiological roles of these enzymes in cartilage will contribute to our understanding of OA pathogenesis and enable design of targeted inhibitors that effectively target metalloproteinase-mediated cartilage degradation while sparing cartilage repair pathways.

  • differential regulation of extracellular tissue inhibitor of metalloproteinases 3 levels by cell membrane bound and shed low density lipoprotein receptor related protein 1
    Journal of Biological Chemistry, 2013
    Co-Authors: Linda Troeberg, Simone D Scilabra, Kazuhiro Yamamoto, Herve Emonard, Ida B Thogersen, Jan J Enghild, Dudley K Strickland, Hideaki Nagase
    Abstract:

    Tissue inhibitor of metalloproteinases-3 (TIMP-3) plays a key role in regulating extracellular matrix turnover by inhibiting matrix metalloproteinases (MMPs), Adamalysins (ADAMs), and Adamalysins with thrombospondin motifs (ADAMTSs). We demonstrate that levels of this physiologically important inhibitor can be regulated post-translationally by endocytosis. TIMP-3 was endocytosed and degraded by a number of cell types including chondrocytes, fibroblasts, and monocytes, and we found that the endocytic receptor low density lipoprotein receptor-related protein-1 (LRP-1) plays a major role in TIMP-3 internalization. However, the cellular uptake of TIMP-3 significantly slowed down after 10 h due to shedding of LRP-1 from the cell surface and formation of soluble LRP-1 (sLRP-1)-TIMP-3 complexes. Addition of TIMP-3 to HTB94 human chondrosarcoma cells increased the release of sLRP-1 fragments of 500, 215, 160, and 110 kDa into the medium in a concentration-dependent manner, and all of these fragments were able to bind to TIMP-3. TIMP-3 bound to sLRP-1, which was resistant to endocytosis, retained its inhibitory activity against metalloproteinases. Extracellular levels of sLRP-1 can thus increase the half-life of TIMP-3 in the extracellular space, controlling the bioavailability of TIMP-3 to inhibit metalloproteinases.

  • microvesicles shed by oligodendroglioma cells and rheumatoid synovial fibroblasts contain aggrecanase activity
    Matrix Biology, 2012
    Co-Authors: Alessandra Lo Cicero, Iwona Majkowska, Italia Di Liegro, Hideaki Nagase, Linda Troeberg
    Abstract:

    Membrane microvesicle shedding is an active process and occurs in viable cells with no signs of apoptosis or necrosis. We report here that microvesicles shed by oligodendroglioma cells contain an ‘aggrecanase’ activity, cleaving aggrecan at sites previously identified as targets for Adamalysin metalloproteinases with disintegrin and thrombospondin domains (ADAMTSs). Degradation was inhibited by EDTA, the metalloproteinase inhibitor GM6001 and by tissue inhibitor of metalloproteinases (TIMP)-3, but not by TIMP-1 or TIMP-2. This inhibitor profile indicates that the shed microvesicles contain aggrecanolytic ADAMTS(s) or related TIMP-3-sensitive metalloproteinase(s). The oligodendroglioma cells were shown to express the three most active aggrecanases, namely Adamts1, Adamts4 and Adamts5, suggesting that one or more of these enzymes may be responsible for the microvesicle activity. Microvesicles shed by rheumatoid synovial fibroblasts similarly degraded aggrecan in a TIMP-3-sensitive manner. Our findings raise the novel possibility that microvesicles may assist oligodendroglioma and rheumatoid synovial fibroblasts to invade through aggrecan-rich extracellular matrices.

  • proteases involved in cartilage matrix degradation in osteoarthritis
    Biochimica et Biophysica Acta, 2012
    Co-Authors: Linda Troeberg, Hideaki Nagase
    Abstract:

    Osteoarthritis is a common joint disease for which there are currently no disease-modifying drugs available. Degradation of the cartilage extracellular matrix is a central feature of the disease and is widely thought to be mediated by proteinases that degrade structural components of the matrix, primarily aggrecan and collagen. Studies on transgenic mice have confirmed the central role of Adamalysin with Thrombospondin Motifs 5 (ADAMTS-5) in aggrecan degradation, and the collagenolytic matrix metalloproteinase MMP-13 in collagen degradation. This review discusses recent advances in current understanding of the mechanisms regulating expression of these key enzymes, as well as reviewing the roles of other proteinases in cartilage destruction. This article is part of a Special Issue entitled: Proteolysis 50 years after the discovery of lysosome.

Didier Cataldo - One of the best experts on this subject based on the ideXlab platform.

  • implication of Adamalysin proteases in mesothelioma
    European Respiratory Journal, 2015
    Co-Authors: Christelle Sepult, Natacha Rocks, Sandrine Bekaert, Kim Donati, Bernard Duysinx, Agnes Noel, Didier Cataldo
    Abstract:

    The main risk factor for malignant pleural mesothelioma (MPM) is asbestos fibers exposure but mechanisms that preclude malignant transformation are not totally elucidated. The implication of ADAM/TS (A Disintegrin And Metalloprotease/with ThromboSpondin motifs) has been previously unveiled in several cancers. We aimed to determine the potential importance of ADAM/TS in mesothelioma progression. We used a mouse model based on orthotopic injection of AB12 cell line (murine malignant mesothelioma cell line) . BALB/C mice were intrapleurally injected with AB12 cells and tumors developed in the pleural space showed an increase of ADAM-8 and -10 expressions as compared to cultured AB12. We also developed a mouse primary cancerization model induced by repeated intraperitoneal injections of crocidolite fibers. After one year, primary mesothelioma cell lines were harvested from these animals. These primary mesothelioma cell lines were injected in pleura of syngeneic animals and displayed an increase of ADAM-8 expression as compared to cultured cells. Twelve human samples of MPM were screened and showed significant increase of ADAM-8 and -10 expressions as compared to matched non-diseased pleural samples. In vitro targeted inhibition studies: showed that ADAM8 depletion decreases MPM cells migration in a wound-healing assay. In conclusion, these results identify ADAM8 as a potential target in mesothelioma.

  • involvement of Adamalysin proteases in modulations of tumor microenvironment and premetastatic niches
    European Respiratory Journal, 2015
    Co-Authors: Kim Donati, Christelle Sepult, Natacha Rocks, Sandrine Bekaert, Agnes Noel, Didier Cataldo
    Abstract:

    According to the “seed and soil” theory, metastases are favored by a permissive microenvironment. Proteases of the Adamalysin family (ADAM/ADAMTS) are modifiers of the biological activity of various mediators and could potentially modulate tumor progression and metastases formation. We aim at determining the involvement of ADAM/ADAMTS at a premetastatic stage in lung parenchyma of mice bearing distant primary tumors. Balb/c mice were subcutaneously injected in each flank with 2.105 Luc+ 4T1 tumor cells and sacrificed at days 3, 7, 9, 14, 21 and 26 after injection. We determined that tumor-bearing mice are in a premetastatic stage at day 7 and already display pulmonary premetastatic niches characteristics (accumulation of LOX, MMP-9 overexpression and crosslinked collagen IV). At day 7, tumor-bearing mice display a neutrophilic inflammation in lung parenchyma. Boyden assays using the bronchoalveolar lavage fluids showed that modulations of the pulmonary microenvironment in tumor-bearing mice induce recruitment of 4T1 cells. We demonstrated an increased ADAM8 expression at the premetastatic stage in the lungs. We conclude that distant primary tumors induce an accumulation of neutrophils and an increase of ADAM8 in the lungs before metastases formation. These changes might increase tumor engraftment in the lung.

  • role of Adamalysin proteases in modulations of tumor microenvironment and premetastatic niches
    Revue Des Maladies Respiratoires, 2015
    Co-Authors: Kim Donati, Christelle Sepult, Natacha Rocks, Sandrine Bekaert, Agnes Noel, Didier Cataldo
    Abstract:

    Introduction Metastasis is the predominant cause of death in patients with cancer and reach relatively frequently specific secondary target organs such as the lungs. According to the “seed and soil” paradigm, these blood-borne metastases can be favored by a permissive microenvironment in these organs promoting tumor cells arrival and metastases growth. Inflammation seems to play a role in the formation of a permissive microenvironment in different organs. The influence of individual inflammatory cell type in the premetastatic niches is less known. Proteases of the Adamalysin family (ADAM and ADAMTS) are implicated both in various physiologic and in pathologic processes such as cancer. As these enzymes are known to be able to induce the cleavage and modification of the biological activity of various biological factors, they could be considered as molecular players that could potentially link the inflammation to tumor progression and metastases formation. Aims During this study, we analyzed the differential expression of ADAM/ADAMTS at a premetastatic stage in the lung parenchyma of mice bearing distant primary tumors. Methods and results Using balb/c mice injected intradermally in each flank with 2 × 105 Luciferase+ 4T1 tumoral cells and sacrified at days 3, 7, 9, 14, 21 and 26 after injection, we report an emergence of detectable metastasis at day 21 by Xenogen IVIS ® and histological lung sections analysis. Tumor-bearing mice quickly display a neutrophilic inflammation (already after 7 days) in lung parenchyma and in the broncho-alveolar lavages. As we intended to assess the potential contribution of Adamalysins in the premetastatic niche, we ensured the absence of tumor cells in the lungs and chosen after different experiments to perform our analysis on the lungs of mice sacrificed on day 7. At day 7, mice already display premetastatic niches characterics such as an accumulation of lysyl oxidase, MMP-9 and crosslinked collagen IV into the lungs. Finally, we showed an increase of ADAM-8 and -17 expression at a premetastatic stage into the lungs of tumour-bearing mice compared to the control mice. Conclusions In this study, we determined the time course of lung metastases for 4T1 cells after intradermal injection. We report that distant primary tumors induce an accumulation of neutrophils and an increase of ADAM-8 and -17 into the lungs before the metastases formation. Perspectives We will determine the pulmonary stromal cells responsible of ADAM-8 and -17 modulations.

Wolfram Bode - One of the best experts on this subject based on the ideXlab platform.

  • the metzincins topological and sequential relations between the astacins Adamalysins serralysins and matrixins collagenases define a superfamily of zinc peptidases
    Protein Science, 2008
    Co-Authors: Walter Stocker, F X Gomisruth, Ulrich Baumann, Frank Grams, Peter Reinemer, David B Mckay, Wolfram Bode
    Abstract:

    The three-dimensional structures of the zinc endopeptidases human neutrophil collagenase, Adamalysin II from rattle snake venom, alkaline proteinase from Pseudomonas aeruginosa, and astacin from crayfish are topologically similar, with respect to a five-stranded beta-sheet and three alpha-helices arranged in typical sequential order. The four proteins exhibit the characteristic consensus motif HEXXHXXGXXH, whose three histidine residues are involved in binding of the catalytically essential zinc ion. Moreover, they all share a conserved methionine residue beneath the active site metal as part of a superimposable "Met-turn." This structural relationship is supported by a sequence alignment performed on the basis of topological equivalence showing faint but distinct sequential similarity. The alkaline proteinase is about equally distant (26% sequence identity) to both human neutrophil collagenase and astacin and a little further away from Adamalysin II (17% identity). The pairs astacin/Adamalysin II, astacin/human neutrophil collagenase, and Adamalysin II/human neutrophil collagenase exhibit sequence identities of 16%, 14%, and 13%, respectively. Therefore, the corresponding four distinct families of zinc peptidases, the astacins, the matrix metalloproteinases (matrixins, collagenases), the Adamalysins/reprolysins (snake venom proteinases/reproductive tract proteins), and the serralysins (large bacterial proteases from Serratia, Erwinia, and Pseudomonas) appear to have originated by divergent evolution from a common ancestor and form a superfamily of proteolytic enzymes for which the designation "metzincins" has been proposed. There is also a faint but significant structural relationship of the metzincins to the thermolysin-like enzymes, which share the truncated zinc-binding motif HEXXH and, moreover, similar topologies in their N-terminal domains.

  • the crystal structure of Adamalysin ii a zinc endopeptidase from the snake venom of the eastern diamondback rattlesnake crotalus adamanteus
    Brazilian Journal of Medical and Biological Research, 1994
    Co-Authors: Wolfram Bode, Lawrence F. Kress, Edgar F Meyer, F X Gomisruth
    Abstract:

    : 1. Adamalysin II, alias proteinase II, a 24-kDa zinc-endopeptidase from the snake venom of Crotalus adamanteus, is a member of a large family of metalloproteinases isolated as small proteinases or proteolytic domains of mosaic hemorrhagic proteins from various snake venoms. Homologous domains have been recently detected in multimodular mammalian reproductive tract proteins and in mammalian gene products, somatic rearrangements of which seem to be linked to primary breast cancers. 2. The 2.0 A X-ray crystal structure of Adamalysin II reveals an ellipsoidal molecule with a shallow active-site cleft separating a relatively irregularly folded sub-domain from the main molecular body composed of a 5-stranded beta-sheet and four alpha-helices. Opposite to this active-site cleft is an integrated calcium ion liganded by carbonyl and strongly conserved carboxylate/carboxamide residues. The folding of the peptide fragment containing the zinc-binding motif HExxHxxGxxH bears only a distant resemblance to thermolysin; it is identical to that found in astacin, in collagenases, and in serralysins, with the three histidines (His142, His146, His152) and a water molecule (linked to the glutamic acid Glu143) likewise constituting the zinc ligand; similar to collagenases, but in contrast to astacin, Adamalysin II lacks a fifth (tyrosine) zinc ligand, leaving its zinc-ion tetrahedrally coordinated. Furthermore, Adamalysin II shares an identical active-site basement formed by a common Met-turn. 3. Due to their virtually identical active-site environment and similar folding topology, the snake venom metalloproteinases (hitherto called Adamalysins) and the three other proteinases might be grouped into a common superfamily called metzincins with distinct differences from the thermolysin family.

  • refined 2 0 a x ray crystal structure of the snake venom zinc endopeptidase Adamalysin ii primary and tertiary structure determination refinement molecular structure and comparison with astacin collagenase and thermolysin
    Journal of Molecular Biology, 1994
    Co-Authors: F X Gomisruth, L F Kress, Josef Kellermann, Irmgard Mayr, Robert Huber, Wolfram Bode
    Abstract:

    Abstract Adamalysin II, alias proteinase II, a 24 kDa zinc-endopeptidase isolated from the snake venom of the Eastern diamondback rattlesnake Crotalus adamanteus , is a prototype, of the proteolytic domain of snake venom metalloproteinases and of domains found in mammalian reproductive tract proteins. Its 2·0 A crystal and molecular structure was solved by multiple isomorphous replacement using six heavy-atom derivatives, and was refined to a crystallographic R -value of 0·172. 201 of the 203 amino acid residues of Adamalysin II are defined by electron density; only the first two residues are disordered and crystallographically undefined in the crystal structure. Three-quarters of these crystallographic amino acid residue assignments were confirmed by chemical sequencing. In addition, the active-site zinc-ion, a heptaco-ordinated calcium ion, a fixed sulphate anion and 173 solvent molecules were localized in the structure. Adamalysin II is an ellipsoidal molecule with a relatively flat active-site cleft separating the "upper" main body from a small "lower" subdomain. The regularly folded N-terminal upper domain consists essentially of a central, highly twisted five-stranded β-pleated sheet flanked by a long and a short surface located helix on its convex side, and by two long helices, one of which represents the central "active site helix", on its concave side. The lower subdomain, comprising the last 50 residues, is organized in multiple turns, with the chain ending in a long C-terminal helix and an extended segment clamped to the upper domain via a disulphide bridge. The catalytic zinc-ion, located at the bottom of the active-site cleft, is almost tetrahedrally co-ordinated by His142, His146 and His152, and a water molecule anchored to an intermediate glutamic acid residue (Glu143), with the three imidazole Ne nitrogen atoms 2·1 A and the solvent oxygen atom 2·4 A away from the zinc ion. His142, Glu143 and His146 are part of the long active-site helix, which extends up to Gly149, where it turns sharply away towards His152. The importance of these residues for structure and activity of Adamalysin II explains their occurrence in the HEXXHXXGXXH consensus sequence. Asp153, which is strictly conserved in these snake venom and reproductive tract metalloproteinases, is buried in the subdomain and seems to stabilize the hydrophobic active-site basement. Some residues behind, the Adamalysin peptide chain folds into a characteristic 1,4-turn (the "Met-turn") containing the conserved Met166, which forms a hydrophobic basement for the three zinc-binding imidazoles. Adamalysin II shares a similar overall topology with astacin, with the collagenase catalytic domains and the P. aeruginosa alkaline proteinase, and exhibits a virtually identical zinc environment with these other "metzincins". The catalytic domains of the snake venom metalloproteinases and the corresponding domains of some reproductive tract proteins can be aligned with only a very few single residue insertions and deletions and with sequence identities mostly above 50%. All substitutions, including some novel cysteine residues engaged in additional disulphide bridges, are in agreement with the Adamalysin 11 structure, which can therefore be considered as a prototype of these "adarnalysins".

Qingxiang Amy Sang - One of the best experts on this subject based on the ideXlab platform.

  • dynamic change of Adamalysin 19 adam19 in human placentas and its effects on cell invasion and adhesion in human trophoblastic cells
    Science China-life Sciences, 2009
    Co-Authors: Qingxiang Amy Sang, Meirong Zhao, Wei Qiu, Yanling Wang
    Abstract:

    Human ADAM19 is a recently identified member of the ADAM family. It is highly expressed in human placentas, but its dynamic change and function at the human feto-maternal interface during placentation remain to be elucidated. In this present study, the spatial and temporal expression and cellular localization of ADAM19 in normal human placentas were first demonstrated, and the effects of ADAM19 on trophoblast cell adhesion and invasion were further investigated by using a human choriocarcinoma cell line (JEG-3) as an in vitro model. The data demonstrated that ADAM19 was widely distributed in villous cytotrophoblast cells, syncytiotrophoblast cells, column trophoblasts, and villous capillary endothelial cells during early pregnancy. The mRNA and protein level of ADAM19 in placentas was high at gestational weeks 8–9, but diminished significantly at mid- and term pregnancy. In JEG-3 cells, the overexpression of ADAM19 led to diminished cell invasion, as well as increases in cell adhesiveness and the expression of E-cadherin, with no changes in β-catenin expression observed. These data indicate that ADAM19 may participate in the coordinated regulation of human trophoblast cell behaviors during the process of placentation.

  • adam19 Adamalysin 19 structure function and role as a putative target in tumors and inflammatory diseases
    Current Pharmaceutical Design, 2009
    Co-Authors: Robert G Newcomer, Qingxiang Amy Sang
    Abstract:

    A disintegrin and metalloproteinase 19 (ADAM19, or Adamalysin 19) is a cell surface glycoprotein with a signal sequence, a prodomain, a metalloproteinase domain, a disintegrin domain, a cysteine-rich domain, a epidermal growth factor-like domain, a transmembrane domain, and a cytoplasmic domain. It is an endopeptidase that cleaves extracellular matrix proteins and sheds growth factors and cytokines such as neuregulins, heparin-binding epidermal growth factor, tumor necrosis factor (TNF)-alpha, and TNF-related activation-induced cytokine. The ADAM19 gene was cloned from human, monkey, and mouse. It is expressed in multiple organs and tissues including heart, lung, bones, brain, spleen, liver, skeletal muscle, kidney, and testes. ADAM19 plays essential roles in embryo implantation, cardiovascular morphogenesis, neurogenesis, and other developmental processes. It has constitutive alpha-secretase activity associated with processing Alzheimer's disease amyloid precursor protein (APP) to non-amyloidogenic fragments; thus, it is neuroprotective. Those observations indicate that inhibition of ADAM19 activity is undesirable during embryo development and morphogenesis, and during the development of Alzheimer's disease. On the contrary, in adults, ADAM19 is upregulated in human brain tumors such as astrocytoma and glioblastoma and is correlated with the invasiveness of glioma. It is also over-expressed by many human cancerous cell lines including cancers of the colon, ovary, lung, and brain. Abnormally high expression of ADAM19 is also linked to inflammation and fibrosis of the lung and kidney. Targeted inhibition of ADAM19 may be crucial for the treatment of certain types of tumors and inflammatory diseases.

  • expression of Adamalysin 19 adam19 in the endometrium and placenta of rhesus monkey macaca mulatta during early pregnancy
    Molecular Human Reproduction, 2005
    Co-Authors: Hongxing Wang, Yunge Zhao, Qingxiang Amy Sang, Hongmei Wang, Qing Yang, Haiyan Lin, Cheng Zhu
    Abstract:

    A disintegrin and metalloproteinase (ADAM) 19 may contribute to multiple processes including proteolysis, adhesion and intracellular signalling. These processes are also critical for embryo implantation. The aim of this study was to investigate the spatio-temporal expression of the ADAM19 in rhesus monkey uteri on days 12, 18 and 26 of pregnancy. The results showed that in the cloned monkey 346 bp ADAM19 gene fragment and 114 amino acid residues were 98 and 100% identical to those of human homologues, respectively. In-situ hybridization confirmed that the ADAM19 mRNA was located in the luminal and glandular epithelium on day 12 of pregnancy. On day 18 of pregnancy, strong signals of the ADAM19 mRNA were detected in the placental villi, trophoblastic column and glandular epithelium near the myometrium. Moderate expression of the ADAM19 mRNA was seen in the trophoblastic shell and stromal cells. The placental villi and trophoblastic column expressed abundant ADAM19 mRNA, and ADAM19 transcripts were also detected in the trophoblastic shell and fetal-maternal border on day 26 of pregnancy. The expression pattern of the ADAM19 protein was similar to its transcript, but signals for the ADAM19 protein in the stromal cells and trophoblastic shell increased more than its mRNA on day 18 of pregnancy. Statistical analysis demonstrated that the expression level of ADAM19 significantly increased on day 18 of pregnancy. These data suggest that the ADAM19 may be involved in the key processes of glandular secretion, trophoblast invasion and degradation of extracellular matrix during early pregnancy.

  • autolytic processing at glu586 ser587 within the cysteine rich domain of human Adamalysin 19 disintegrin metalloproteinase 19 is necessary for its proteolytic activity
    Journal of Biological Chemistry, 2002
    Co-Authors: Yunge Zhao, Tiebang Kang, Hyun I Park, Yewseok Suh, Harald Tschesche, Qingxiang Amy Sang
    Abstract:

    We investigated the regulation of the proteolytic activity of human Adamalysin 19 (a disintegrin and metalloproteinase 19, hADAM19). It was processed at Glu(586)(P1)-Ser(587)(P1') site in the cysteine-rich domain as shown by protein N-terminal sequencing. This truncation was autolytic as illustrated by its R199A/R200A or E346A mutation that prevented the zymogen activation by furin or abolished the catalytic activity. Reagents that block furin-mediated activation of pro-hADAM19, decRVKR-CMK, and brefeldin A abrogated this processing. The sizes of the side chains of the P1 and P1' residues are critical for the processing of hADAM19. The amount of processing product in the E586Q or S587A mutant with a side chain almost the same size as that in the wild type was almost equal. Conversely, very little processing was observed when the size of the side chain was changed significantly, such as in the E586A, E586G, or S587F mutants. Two mutants with presumably subtle structural distinctions from wild type hADAM19, E586D and S587T, displayed rare or little processing and had very low capacities to cleave alpha2-macroglobulin and a peptide substrate. Therefore, this processing is necessary for hADAM19 to exert its proteolytic activities. Moreover, a new peptide substrate, Ac-RPLE-SNAV, which is identical to the processing site sequence, was cleaved at the E-S bond by soluble hADAM19 containing the catalytic and disintegrin domains. This enzyme cleaved the substrate with K(m), k(cat), and k(cat)/K(m) of 2.0 mm, 2.4/min, and 1200 m(-1) min(-1), respectively, using a fluorescamine assay. Preliminary studies showed that a protein kinase C activator, phorbol 12-myristate 13-acetate, promoted the cellular processing of hADAM19; however, three calmodulin antagonists, trifluoperazine, W7, and calmidazolium, impaired this cleavage, indicating complex signal pathways may be involved in the processing.

  • intracellular activation of human Adamalysin 19 disintegrin and metalloproteinase 19 by furin occurs via one of the two consecutive recognition sites
    Journal of Biological Chemistry, 2002
    Co-Authors: Tiebang Kang, Yunge Zhao, Duanqing Pei, Joseph F Sucic, Qingxiang Amy Sang
    Abstract:

    Abstract Adamalysin 19 (a disintegrin and metalloproteinase 19, ADAM19, or meltrin β) is a plasma membrane metalloproteinase. Human ADAM19 zymogen contains two potential furin recognition sites (RX(K/R)R),196KRPR 200 Rand199RRMK 203 R, between its pro- and catalytic domains. Protein N-terminal sequencing revealed that the cellular mature forms of hADAM19 started at204EDLNSMK, demonstrating that the preferred furin cleavage site was the200 RMK 203 R↓204EDLN. Those mature forms were catalytically active. Both Pittsburgh mutant of α1-proteinase inhibitor and dec-Arg-Val-Lys-Arg-chloromethyl ketone, two specific furin inhibitors, blocked the activation of hADAM19. Activation of hADAM19 was also blocked by brefeldin A, which inhibits protein trafficking from the endoplasmic reticulum to the Golgi, or A23187, a calcium ionophore known to inhibit the autoactivation of furin. When 202KR were mutated to AA, the proenzyme was also activated, suggesting that197 RPRR is an alternative activation site. Furthermore, only pro-forms of hADAM19 were detected in the 199RR to AA mutant, which abolished both furin recognition sites. Moreover, the zymogens were not converted into their active forms in two furin-deficient mammalian cell lines; co-expression of hADAM19 and furin in these two cell lines restored zymogen activation. Finally, co-localization between furin and hADAM19 was identified in the endoplasmic reticulum-Golgi complex and/or the trans-Golgi network. This report is the first thorough investigation of the intracellular activation of Adamalysin 19, demonstrating that furin activated pro-hADAM19 in the secretory pathway via one of the two consecutive furin recognition sites.