ADAM17 - Explore the Science & Experts | ideXlab

Scan Science and Technology

Contact Leading Edge Experts & Companies

ADAM17

The Experts below are selected from a list of 129669 Experts worldwide ranked by ideXlab platform

ADAM17 – Free Register to Access Experts & Abstracts

Carl P. Blobel – One of the best experts on this subject based on the ideXlab platform.

  • Role of ADAM17 in endotoxin-induced pulmonary inflammation
    European Respiratory Journal, 2011
    Co-Authors: Daniela Dreymueller, Carl P. Blobel, Franz M. Hess, Christian Martin, Tanja Kogel, Keisuke Horiuchi, Stefan Uhlig, Andreas Ludwig
    Abstract:

    Acute lung inflammation is associated with enhanced vascular permeability and leukocyte recruitment. Several proinflammatory, soluble and surface-expressed mediators, including TNFα, TNFR1/2, amphiregulin, IL-6R, IL-1R, L-selectin, CX3CL1, and JAMs may become released by the activity of the metalloproteinases ADAM10 and ADAM17. We examined the role of these proteases in vascular permeability and leukocyte transmigration in vitro by pharmacological inhibition and lentiviral-mediated siRNA knockdown of ADAM10 and ADAM17. The in vivo role of these proteases was studied in a murine model of LPS-induced lung injury by pharmacological inhibition. The relevance of ADAM17 was further analyzed by knockout of ADAM17 in endothelial or smooth muscle cells. The BAL protein levels and the wet/dry-ratio served as markers of vascular permeability and edema formation. Cell recruitment to the alveolar space and lung tissue was analyzed by flow cytometry; cytokines were determined by ELISA. In vitro, transmigration of neutrophils to IL-8 through pulmonary endothelial cells was reduced by pharmacological inhibition as well as knockdown of ADAM10 or ADAM17. LPS-mediated induction of permeability was reduced by pharmacological inhibition, but not by ADAM10 knockdown, indicating a predominant role of ADAM17 in the regulation of endothelial permeability. In vivo, LPS challenge increased the wet/dry-ratio as well as the BAL levels of protein, TNFα, IL-6 and leukocytes. All these effects were largely prevented by inhibitor application or by knockout of ADAM17 in smooth muscle or endothelial cells. These results indicate that local ADAM17 is involved in the onset of inflammation and tissue injury during endotoxin-induced lung inflammation.

  • ADAM17 is regulated by a rapid and reversible mechanism that controls access to its catalytic site
    Journal of Cell Science, 2010
    Co-Authors: Sylvain Le Gall, Paul Saftig, Karina Reiss, Thorsten Maretzky, Priya D Issuree, Xiaoda Niu, Rama Khokha, Daniel Lundell, Carl P. Blobel
    Abstract:

    Protein ectodomain shedding is crucial for cell–cell interactions because it controls the bioavailability of soluble tumor necrosis factor-α (TNFα) and ligands of the epidermal growth factor (EGF) receptor, and the release of many other membrane proteins. Various stimuli can rapidly trigger ectodomain shedding, yet much remains to be learned about the identity of the enzymes that respond to these stimuli and the mechanisms underlying their activation. Here, we demonstrate that the membrane-anchored metalloproteinase ADAM17, but not ADAM10, is the sheddase that rapidly responds to the physiological signaling pathways stimulated by thrombin, EGF, lysophosphatidic acid and TNFα. Stimulation of ADAM17 is swift and quickly reversible, and does not depend on removal of its inhibitory pro-domain by pro-protein convertases, or on dissociation of an endogenous inhibitor, TIMP3. Moreover, activation of ADAM17 by physiological stimuli requires its transmembrane domain, but not its cytoplasmic domain, arguing against inside–out signaling via cytoplasmic phosphorylation as the underlying mechanism. Finally, experiments with the tight binding hydroxamate inhibitor DPC333, used here to probe the accessibility of the active site of ADAM17, demonstrate that this inhibitor can quickly bind to ADAM17 in stimulated, but not quiescent cells. These findings support the concept that activation of ADAM17 involves a rapid and reversible exposure of its catalytic site.

  • stimulation of platelet derived growth factor receptor β pdgfrβ activates ADAM17 and promotes metalloproteinase dependent cross talk between the pdgfrβ and epidermal growth factor receptor egfr signaling pathways
    Journal of Biological Chemistry, 2010
    Co-Authors: Karen Mendelson, Paul Saftig, Carl P. Blobel, Steven Swendeman
    Abstract:

    Binding of the platelet-derived growth factor (PDGF)-B to its receptor PDGFRβ promotes proliferation, migration, and recruitment of pericytes and smooth muscle cells to endothelial cells, serving to stabilize developing blood vessels. The main goals of this study were to determine whether the extracellular domain of the PDGFRβ can be proteolytically released from cell membranes and, if so, to identify the responsible sheddase and determine whether activation of the PDGFRβ stimulates its shedding and potentially that of other membrane proteins. We found that the PDGFRβ is shed from cells by a metalloproteinase and used loss-of-function experiments to identify ADAM10 as the sheddase responsible for constitutive and ionomycin-stimulated processing of the PDGFRβ. Moreover, we showed that ligand-dependent activation of the PDGFRβ does not trigger its own shedding by ADAM10, but instead it stimulates ADAM17 and shedding of substrates of ADAM17, including tumor necrosis factor α and transforming growth factor α. Finally, we demonstrated that treatment of mouse embryonic fibroblasts with PDGF-B triggers a metalloproteinase-dependent cross-talk between the PDGFRβ and the epidermal growth factor receptor (EGFR)/ERK1/2 signaling axis that is also critical for PDGF-B-stimulated cell migration, most likely via ADAM17-dependent release and activation of ligands of the EGFR. This study identifies the principal sheddase for the PDGFRβ and provides new insights into the mechanism of PDGFRβ-dependent signal transduction and cross-talk with the EGFR.

Paul Saftig – One of the best experts on this subject based on the ideXlab platform.

  • 230: Unraveling the role of ADAM17 in IL-6 trans-signaling
    Cytokine, 2013
    Co-Authors: Jeanette Schwarz, Paul Saftig, Isabell Yan, Olga Will, Stefan-rose-john, Hans-willi Mittrücker, Athena Chalaris
    Abstract:

    IL-6 is a key regulator of immune responses after bacterial infection. IL-6 signaling is mediated via the receptor subunits IL-6R and gp130. The signal transducing subunit gp130 is ubiquitously expressed whereas IL-6R expression is restricted to hepatocytes and some leukocytes. The IL-6R can be cleaved from the cell surface by ADAM proteases. The soluble form of the IL-6R (sIL-6R) has the same IL-6 binding affinity as the membrane-bound receptor. The resulting IL-6/sIL-6R complex activates cells, which only express gp130 on their cell surface, a process called trans-signaling. Thus, IL-6-transsignaling following IL-6R proteolysis essentially renders all cells of the body responsive to IL-6. The proteolytic web controlling IL-6R shedding in vivo under pathophysiological conditions is poorly characterized. Recently, it was demonstrated that human IL-6R is a substrate for the protease ADAM17 in contrast to the murine ADAM17, which is mainly cleaved by ADAM10. However, these data were raised in cell culture systems. We analyzed IL-6R shedding in vivo on leukocytes after infection with Listeria monocytogenes or LPS-challenge of ADAM17 and ADAM10-deficient mice. ADAM17 and ADAM10 knockout mice are not viable. For this reason we generated a hypomorphic ADAM17 mouse model which expresses only 5% of the normal ADAM17 levels. Additionally we use conditional ADAM10 mice with ADAM10 deleted either in monocytes or Tcells. Listeria infection led to massive shedding of the IL-6R on Tcells and inflammatory monocytes. Further we could demonstrate that Listeria -induced IL-6R cleavage is abrogated in hypomorphic ADAM17 ex/ex mice. Additionally we can show that serum levels of sIL-6R are elevated after LPS challenge of wildtype but not ADAM17 ex/ex mice. These results strongly suggest that ADAM17 is the main IL-6R sheddase under pathophysiological conditions. In future experiments we will evaluate the contribution of the protease ADAM10 in mediating IL-6R shedding to uncover the proteolytic steps involved in IL-6 trans-signaling.

  • the a disintegrin and metalloproteases adam10 and ADAM17 novel drug targets with therapeutic potential
    European Journal of Cell Biology, 2011
    Co-Authors: Paul Saftig, Karina Reiss
    Abstract:

    Proteolytic ectodomain release, a process known as “shedding”, has been recognised as a key mechanism for regulating the function of a diversity of cell surface proteins. A Disintegrin And Metalloproteinases (ADAMs) have emerged as the major proteinase family that mediates ectodomain shedding. Dysregulation of ectodomain shedding is associated with autoimmune and cardiovascular diseases, neurodegeneration, infection, inflammation and cancer. Therefore, ADAMs are increasingly regarded as attractive targets for novel therapies. ADAM10 and its close relative ADAM17 (TNF-alpha converting enzyme (TACE)) have been studied in particular in the context of ectodomain shedding and have been demonstrated as key molecules in most of the shedding events characterised to date. Whereas the level of expression of ADAM10 may be of importance in cancer and neurodegenerative disorders, ADAM17 mainly coordinates pro- and anti-inflammatory activities during immune response. Despite the high therapeutical potential of ADAM inhibition, all clinical trials using broad-spectrum metalloprotease inhibitors have failed so far. This review will cover the emerging roles of both ADAM10 and ADAM17 in the regulation of major physiological and developmental pathways and will discuss the suitability of specifically modulating the activities of both proteases as a feasible way to inhibit inflammatory states, cancer and neurodegeneration.

  • ADAM17 is regulated by a rapid and reversible mechanism that controls access to its catalytic site
    Journal of Cell Science, 2010
    Co-Authors: Sylvain Le Gall, Paul Saftig, Karina Reiss, Thorsten Maretzky, Priya D Issuree, Xiaoda Niu, Rama Khokha, Daniel Lundell, Carl P. Blobel
    Abstract:

    Protein ectodomain shedding is crucial for cell–cell interactions because it controls the bioavailability of soluble tumor necrosis factor-α (TNFα) and ligands of the epidermal growth factor (EGF) receptor, and the release of many other membrane proteins. Various stimuli can rapidly trigger ectodomain shedding, yet much remains to be learned about the identity of the enzymes that respond to these stimuli and the mechanisms underlying their activation. Here, we demonstrate that the membrane-anchored metalloproteinase ADAM17, but not ADAM10, is the sheddase that rapidly responds to the physiological signaling pathways stimulated by thrombin, EGF, lysophosphatidic acid and TNFα. Stimulation of ADAM17 is swift and quickly reversible, and does not depend on removal of its inhibitory pro-domain by pro-protein convertases, or on dissociation of an endogenous inhibitor, TIMP3. Moreover, activation of ADAM17 by physiological stimuli requires its transmembrane domain, but not its cytoplasmic domain, arguing against inside–out signaling via cytoplasmic phosphorylation as the underlying mechanism. Finally, experiments with the tight binding hydroxamate inhibitor DPC333, used here to probe the accessibility of the active site of ADAM17, demonstrate that this inhibitor can quickly bind to ADAM17 in stimulated, but not quiescent cells. These findings support the concept that activation of ADAM17 involves a rapid and reversible exposure of its catalytic site.

Jurgen Scheller – One of the best experts on this subject based on the ideXlab platform.

  • human and murine interleukin 23 receptors are novel substrates for a disintegrin and metalloproteases adam10 and ADAM17
    Journal of Biological Chemistry, 2016
    Co-Authors: Manuel Franke, Christoph Garbers, Doreen M Floss, Christoph Beckerpauly, Bjorn Rabe, Jutta Schroder, Niloufar Monhasery, Theresa Ackfeld, Thorben M Hummel, Jurgen Scheller
    Abstract:

    IL-23 (interleukin 23) regulates immune responses against pathogens and plays a major role in the differentiation and maintenance of TH17 cells and the development of autoimmune diseases and cancer. The IL-23 receptor (IL-23R) complex consists of the unique IL-23R and the common IL-12 receptor β1 (IL-12Rβ1). Differential splicing generates antagonistic soluble IL-23R (sIL-23R) variants, which might limit IL-23-mediated immune responses. Here, ectodomain shedding of human and murine IL-23R was identified as an alternative pathway for the generation of sIL-23R. Importantly, proteolytically released sIL-23R has IL-23 binding activity. Shedding of IL-23R was induced by stimulation with the phorbol ester phorbol 12-myristate 13-acetate (PMA), but not by ionomycin. PMA-induced shedding was abrogated by an ADAM (A disintegrin and metalloprotease) 10 and 17 selective inhibitor, but not by an ADAM10 selective inhibitor. ADAM17-deficient but not ADAM10-deficient HEK293 cells failed to shed IL-23R after PMA stimulation, demonstrating that ADAM17 but not ADAM10 cleaves the IL-23R. Constitutive shedding was, however, inhibited by an ADAM10 selective inhibitor. Using deletions and specific amino acid residue exchanges, we identified critical determinants of ectodomain shedding within the stalk region of the IL-23R. Finally, interaction studies identified domains 1 and 3 of the IL-23R as the main ADAM17 binding sites. In summary, we describe human and murine IL-23R as novel targets for protein ectodomain shedding by ADAM10 and ADAM17.

  • t cell immunoglobulin and mucin domain 2 tim 2 is a target of adam10 mediated ectodomain shedding
    FEBS Journal, 2014
    Co-Authors: Christin Dewitz, Jurgen Scheller, Athena Chalaris, Karina Reiss, Katja Mollerhackbarth, Olga Schweigert, Stefan Rosejohn
    Abstract:

    T-cell immunoglobulin and mucin domain (TIM)-2 is expressed on activated B cells. Here, we provide evidence that murine TIM-2 is a target of ADAM10-mediated ectodomain shedding, resulting in the generation of a soluble form of TIM-2. We identified ADAM10 but not ADAM17 as the major sheddase of TIM-2, as shown by pharmacological ADAM10 inhibition and with ADAM10-deficient and ADAM17-deficient murine embryonic fibroblasts. Ionomycin-induced or 2′(3′)-O-(4-benzoylbenzoyl) ATP triethylammonium salt-induced shedding of TIM-2 was abrogated by deletion of 10 juxtamembrane amino acids from the stalk region but not by deletion of two further N-terminally located blocks of 10 amino acids, indicating a membrane-proximal cleavage site. TIM-2 lacking the intracellular domain was cleaved after ionomycin or 2′ (3′)-O-(4-benzoylbenzoyl) ATP triethylammonium salt treatment, indicating that this domain was not involved in the regulation of ectodomain shedding. Moreover, TIM-2 shedding was negatively controlled by calmodulin. Shed and soluble TIM-2 interacted with H-ferritin. In summary, we describe TIM-2 as a novel target for ADAM10-mediated ectodomain shedding, and reveal the involvement of ADAM proteases in cellular iron homeostasis.

  • a disintegrin and metalloprotease adam 10 and ADAM17 are major sheddases of t cell immunoglobulin and mucin domain 3 tim 3
    Journal of Biological Chemistry, 2013
    Co-Authors: Katja Mollerhackbarth, Christoph Garbers, Stefan Rosejohn, Christin Dewitz, Olga Schweigert, Ahmad Trad, Jurgen Scheller
    Abstract:

    T cell immunoglobulin and mucin domain 3 (Tim-3) dampens the response of CD4+ and CD8+ effector T cells via induction of cell death and/or T cell exhaustion and enhances the ability of macrophages to clear pathogens via binding to galectin 9. Here we provide evidence that human Tim-3 is a target of A disintegrin and metalloprotease (ADAM)-mediated ectodomain shedding resulting in a soluble form of Tim-3. We identified ADAM10 and ADAM17 as major sheddases of Tim-3 as shown by ADAM-specific inhibitors and the ADAM10 pro-domain in HEK293 cells and ADAM10/ADAM17-deficient murine embryonic fibroblasts. PMA-induced shedding of Tim-3 was abrogated by deletion of amino acids Glu181–Asp190 of the stalk region and Tim-3 lacking the intracellular domain was not efficiently cleaved after PMA stimulation. Surprisingly, a single lysine residue within the intracellular domain rescues shedding of Tim-3. Shedding of endogenous Tim-3 was found in primary human CD14+ monocytes after PMA and ionomycin stimulation. Importantly, the recently described down-regulation of Tim-3 from Toll-like receptor-activated CD14+ monocytes was caused by ADAM10- and ADAM17-mediated shedding. Inhibition of Tim-3 shedding from lipopolysaccharide-induced monocytes did not influence lipopolysaccharide-induced TNFα and IL-6 but increases IL-12 expression. In summary, we describe Tim-3 as novel target for ADAM-mediated ectodomain shedding and suggest a role of Tim-3 shedding in TLR-mediated immune responses of CD14+ monocytes.