DOCSLIB.ORG

Receptor Mediated Catabolism of Plasminogen Activators

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Receptor Mediated Catabolism of Plasminogen Activators RECEPTOR MEDIATED CATABOLISM OF PLASMINOGEN ACTIVATORS by Philip George Grimsley Thesis submitted for the degree of Doctor of Philosophy from The University of New South Wales School of Medical Sciences 2009 PLEASE TYPE THE UNIVERSITY OF NEW SOUTH WALES Thesis/Dissertation Sheet Surname or Family name: Grimsley First name: Philip Other name/s: George Abbreviation for degree as given in the University calendar: PhD School: School of Medical Sciences Faculty: Medicine Title: Receptor Mediated Catabolism of Plasminogen Activators Abstract 350 words maximum: (PLEASE TYPE) Humans have two plasminogen activators (PAs), tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA), which generate plasmin to breakdown fibrin and other barriers to cell migration. Both PAs are used as pharmaceuticals but their efficacies are limited by their rapid clearance from the circulation, predominantly by parenchymal cells of the liver. At the commencement of the work presented here, the hepatic receptors responsible for mediating the catabolism of the PAs were little understood. tPA degradation by hepatic cell lines was known to depend on the formation of binary complexes with the major PA inhibitor, plasminogen activator inhibitor type-1 (PAI-1). Initial studies presented here established that uPA was catabolised in a fashion similar to tPA by the hepatoma cell line, HepG2. Other laboratories around this time found that the major receptor mediating the binding and endocytosis of the PAs is Low Density Lipoprotein Receptor-related Protein (LRP1). LRP1 is a giant 600 kDa protein that binds a range of structurally and functionally diverse ligands including, activated α2-macroglobulin, apolipoproteins, β-amyloid precursor protein, and a number of serpin-enzymes complexes, including PA•PAI-1 complexes. Further studies for the work presented here centred on this receptor. By using radiolabelled binding assays, ligand blots, and Western blots on cultured cells, the major findings are that: (1) basal LRP1 expression on HepG2 is low compared to a clone termed, HepG2a16, but appears to increase in long term culture; (2) a soluble form of LRP1, which retains ligand-binding capacity, is present in human circulation; (3) soluble LRP1 is also present in cerebral spinal fluid where its role in neurological disorders such as Alzheimer‟s disease is a developing area of interest; and (4) the release of LRP1 is a mechanism conserved in evolution, possibly as distantly as molluscs. The discovery, identification, and characterisation of soluble LRP1 introduces this protein in the human circulation, and presents a possible further level of regulation for its associated receptor system. Declaration relating to disposition of project thesis/dissertation I hereby grant to the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or in part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all property rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstracts International (this is applicable to doctoral theses only). …………………………………………………………… ……………………………………..……………… ……….……………………... Signature Witness ……. Date The University recognises that there may be exceptional circumstances requiring restrictions on copying or conditions on use. Requests for restriction for a period of up to 2 years must be made in writing. Requests for a longer period of restriction may be considered in exceptional circumstances and require the approval of the Dean of Graduate Research. FOR OFFICE USE ONLY Date of completion of requirements for Award: THIS SHEET IS TO BE GLUED TO THE INSIDE FRONT COVER OF THE THESIS To Leonie COPYRIGHT STATEMENT ‘I hereby grant the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstract International (this is applicable to doctoral theses only). I have either used no substantial portions of copyright material in my thesis or I have obtained permission to use copyright material; where permission has not been granted I have applied/will apply for a partial restriction of the digital copy of my thesis or dissertation.' Signed ……………………………………………........................... Date ……………………………………………........................... AUTHENTICITY STATEMENT ‘I certify that the Library deposit digital copy is a direct equivalent of the final officially approved version of my thesis. No emendation of content has occurred and if there are any minor variations in formatting, they are the result of the conversion to digital format.’ Signed ……………………………………………........................... Date ……………………………………………........................... i ORIGINALITY STATEMENT „I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.‟ Signed …………………………………………….............. Date …………………………………………….............. ii PUBLICATIONS Philip G Grimsley, Kathryn A Quinn, Colin N Chesterman, Dwain A Owensby “Evolutionary conservation of circulating low density lipoprotein receptor-related protein-like („LRP1-like‟) molecules” Thrombosis Research 1999; 94(3): 153-164 Philip G Grimsley, Kathryn A Quinn, Dwain A Owensby “Soluble Low density lipoprotein receptor related protein” Trends in Cardiovascular Medicine 1998; 8(8): 363-368 Philip G Grimsley, Kathryn A Quinn, Colin N Chesterman, Dwain A Owensby “Low density lipoprotein receptor-related protein (LRP1) expression varies among HepG2 cell lines” Thrombosis Research 1997; 88(6): 485-98 Kathryn A Quinn, Philip G Grimsley, Yang-Ping Dai, Michael Tapner, Colin N Chesterman, Dwain A Owensby “Soluble low density lipoprotein receptor-related protein (LRP1) circulates in human plasma” Journal of Biological Chemistry 1997; 272(38): 23946-23951 Philip G Grimsley, John F Normyle, Ruth A Brandt, Georgina Joulianos, Colin N Chesterman, Philip J Hogg, Dwain A Owensby “Urokinase binding and catabolism by HepG2 cells is plasminogen activator inhibitor-1 dependent, analogous to interactions of tissue-type plasminogen activator with these cells” Thrombosis Research 1995; 79(4): 353-361 iii CONFERENCE PRESENTATIONS Philip G Grimsley, Kathryn A Quinn, Colin N Chesterman, Dwain A Owensby “Soluble low density lipoprotein receptor-related protein (LRP1) is present in the serum of mammals and birds” Australian Vascular Biology Society Fifth annual scientific meeting, The Fairmont Resort, Leura, Blue Mountains, NSW, 11-13 Sep 1997, page 65 Philip G Grimsley, Kathryn A Quinn, Colin N Chesterman, Dwain A Owensby “A soluble form of low density lipoprotein receptor-related protein (LRP1) circulates in plasma” Australian Vascular Biology Society Fourth annual scientific meeting, Marysville, Victoria, 17-20 Oct 1996, page 43 Philip G Grimsley, Kathryn A Quinn, Colin N Chesterman, Dwain A Owensby “Low density lipoprotein receptor-related protein exists as a soluble form in plasma” Australian Society for Experimental Pathology 28th annual meeting, University of NSW, Sydney, 2-4 Oct 1996, page 18 Philip G Grimsley, Kathryn A Quinn, Philip J Hogg, Colin N Chesterman, Dwain A Owensby “Variation in the level of expression of low density lipoprotein receptor related protein (LRP1) between sublines of HepG2 cells” Australian Vascular Biology Society Third annual conference, Terrigal, NSW, 5-7 Oct 1995 Philip G Grimsley, Fei Le, Philip J Hogg, Colin N Chesterman, Dwain A Owensby “Evidence for a receptor for plasminogen activatorplasminogen activator inhibitor type-1 complexes (PAPAI-1) distinct from the low density lipoprotein receptor-related protein (LRP1) on HepG2 cells” Australian Vascular Biology Society Second annual scientific meeting, Hahndorf, SA, 17-20 Nov 1994 P.G.Grimsley, J.F.Normyle, G.Joulianos, P.J.Hogg, C.N.Chesterman, D.A.Owensby “Specificity of the receptor for tissue type plasminogen activator-plasminogen activator inhibitor type-1 complex (tPAPAI-1) on human hepatoma cell line HepG2 extends to analogous complexes with urokinase (uPAPAI-1)” Australian Vascular Biology Society First scientific meeting, Oasis Resort, Caloundra, Qld, 19-21 Aug 1993, page 42 iv TABLE OF CONTENTS Thesis / Dissertation Sheet Cover Page Dedication Copyright Statement ..................................................................................................................
Load more

Recommended publications
  • The Role of Streptococcal and Staphylococcal Exotoxins and Proteases in Human Necrotizing Soft Tissue Infections
    toxins Review The Role of Streptococcal and Staphylococcal Exotoxins and Proteases in Human Necrotizing Soft Tissue Infections Patience Shumba 1, Srikanth Mairpady Shambat 2 and Nikolai Siemens 1,* 1 Center for Functional Genomics of Microbes, Department of Molecular Genetics and Infection Biology, University of Greifswald, D-17489 Greifswald, Germany; [email protected] 2 Division of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, University of Zurich, CH-8091 Zurich, Switzerland; [email protected] * Correspondence: [email protected]; Tel.: +49-3834-420-5711 Received: 20 May 2019; Accepted: 10 June 2019; Published: 11 June 2019 Abstract: Necrotizing soft tissue infections (NSTIs) are critical clinical conditions characterized by extensive necrosis of any layer of the soft tissue and systemic toxicity. Group A streptococci (GAS) and Staphylococcus aureus are two major pathogens associated with monomicrobial NSTIs. In the tissue environment, both Gram-positive bacteria secrete a variety of molecules, including pore-forming exotoxins, superantigens, and proteases with cytolytic and immunomodulatory functions. The present review summarizes the current knowledge about streptococcal and staphylococcal toxins in NSTIs with a special focus on their contribution to disease progression, tissue pathology, and immune evasion strategies. Keywords: Streptococcus pyogenes; group A streptococcus; Staphylococcus aureus; skin infections; necrotizing soft tissue infections; pore-forming toxins; superantigens; immunomodulatory proteases; immune responses Key Contribution: Group A streptococcal and Staphylococcus aureus toxins manipulate host physiological and immunological responses to promote disease severity and progression. 1. Introduction Necrotizing soft tissue infections (NSTIs) are rare and represent a more severe rapidly progressing form of soft tissue infections that account for significant morbidity and mortality [1].
    [Show full text]
  • Role of Vegetation-Associated Protease Activity in Valve Destruction in Human Infective Endocarditis
    Role of Vegetation-Associated Protease Activity in Valve Destruction in Human Infective Endocarditis Ghada Al-Salih1, Nawwar Al-Attar2,3, Sandrine Delbosc1, Liliane Louedec1, Elisabeth Corvazier1, Ste´phane Loyau1, Jean-Baptiste Michel1,3, Dominique Pidard1,4, Xavier Duval5, Olivier Meilhac1,3,6* 1 INSERM U698, Paris, France, 2 Cardiovascular Surgery Department, Bichat Hospital, AP-HP, Paris, France, 3 University Paris Diderot, Sorbonne Paris Cite´, Paris, France, 4 Institut National des Sciences du Vivant, Centre National de la Recherche Scientifique, Paris, France, 5 INSERM CIC 007, Paris, France, 6 Bichat Stroke Centre, AP-HP, Paris, France Abstract Aims: Infective endocarditis (IE) is characterized by septic thrombi (vegetations) attached on heart valves, consisting of microbial colonization of the valvular endocardium, that may eventually lead to congestive heart failure or stroke subsequent to systemic embolism. We hypothesized that host defense activation may be directly involved in tissue proteolytic aggression, in addition to pathogenic effects of bacterial colonization. Methods and Results: IE valve samples collected during surgery (n = 39) were dissected macroscopically by separating vegetations (VG) and the surrounding damaged part of the valve from the adjacent, apparently normal (N) valvular tissue. Corresponding conditioned media were prepared separately by incubation in culture medium. Histological analysis showed an accumulation of platelets and polymorphonuclear neutrophils (PMNs) at the interface between the VG and the underlying tissue. Apoptotic cells (PMNs and valvular cells) were abundantly detected in this area. Plasminogen activators (PA), including urokinase (uPA) and tissue (tPA) types were also associated with the VG. Secreted matrix metalloproteinase (MMP) 9 was also increased in VG, as was leukocyte elastase and myeloperoxidase (MPO).
    [Show full text]
  • Effects of Glycosylation on the Enzymatic Activity and Mechanisms of Proteases
    International Journal of Molecular Sciences Review Effects of Glycosylation on the Enzymatic Activity and Mechanisms of Proteases Peter Goettig Structural Biology Group, Faculty of Molecular Biology, University of Salzburg, Billrothstrasse 11, 5020 Salzburg, Austria; [email protected]; Tel.: +43-662-8044-7283; Fax: +43-662-8044-7209 Academic Editor: Cheorl-Ho Kim Received: 30 July 2016; Accepted: 10 November 2016; Published: 25 November 2016 Abstract: Posttranslational modifications are an important feature of most proteases in higher organisms, such as the conversion of inactive zymogens into active proteases. To date, little information is available on the role of glycosylation and functional implications for secreted proteases. Besides a stabilizing effect and protection against proteolysis, several proteases show a significant influence of glycosylation on the catalytic activity. Glycans can alter the substrate recognition, the specificity and binding affinity, as well as the turnover rates. However, there is currently no known general pattern, since glycosylation can have both stimulating and inhibiting effects on activity. Thus, a comparative analysis of individual cases with sufficient enzyme kinetic and structural data is a first approach to describe mechanistic principles that govern the effects of glycosylation on the function of proteases. The understanding of glycan functions becomes highly significant in proteomic and glycomic studies, which demonstrated that cancer-associated proteases, such as kallikrein-related peptidase 3, exhibit strongly altered glycosylation patterns in pathological cases. Such findings can contribute to a variety of future biomedical applications. Keywords: secreted protease; sequon; N-glycosylation; O-glycosylation; core glycan; enzyme kinetics; substrate recognition; flexible loops; Michaelis constant; turnover number 1.
    [Show full text]
  • Highly Potent and Selective Plasmin Inhibitors Based on the Sunflower Trypsin Inhibitor-1 Scaffold Attenuate Fibrinolysis in Plasma
    Highly Potent and Selective Plasmin Inhibitors Based on the Sunflower Trypsin Inhibitor-1 Scaffold Attenuate Fibrinolysis in Plasma Joakim E. Swedberg,‡† Guojie Wu,§† Tunjung Mahatmanto,‡# Thomas Durek,‡ Tom T. Caradoc-Davies,∥ James C. Whisstock,§* Ruby H.P. Law§* and David J. Craik‡* ‡Institute for Molecular Bioscience, The University of Queensland, Brisbane QLD 4072, Australia §ARC Centre of Excellence in Advanced Molecular Imaging, Department of Biochemistry and Molecular Biology, Biomedical Discovery Institute, Monash University, VIC 3800, Australia. ∥Australian Synchrotron, 800 Blackburn Road, Clayton, Melbourne, VIC 3168, Australia. †J.E.S. and G.W. contributed equally to this work. Keywords: Antifibrinolytics; Fibrinolysis; Inhibitors; Peptides; Plasmin ABSTRACT Antifibrinolytic drugs provide important pharmacological interventions to reduce morbidity and mortality from excessive bleeding during surgery and after trauma. Current drugs used for inhibiting the dissolution of fibrin, the main structural component of blood clots, are associated with adverse events due to lack of potency, high doses and non-selective inhibition mechanisms. These deficiencies warrant the development of a new generation highly potent and selective fibrinolysis inhibitors. Here we use the 14-amino acid backbone-cyclic sunflower trypsin inhibitor-1 scaffold to design a highly potent (Ki = 0.05 nM) inhibitor of the primary serine protease in fibrinolysis, plasmin. This compound displays a million-fold selectivity over other serine proteases in blood, inhibits fibrinolysis in plasma more effectively than the gold-standard therapeutic inhibitor aprotinin and is a promising candidate for development of highly specific fibrinolysis inhibitors with reduced side effects. 1 INTRODUCTION The physiological process of fibrinolysis regulates the dissolution of blood clots and thrombosis.
    [Show full text]
  • The Plasmin–Antiplasmin System: Structural and Functional Aspects
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Bern Open Repository and Information System (BORIS) Cell. Mol. Life Sci. (2011) 68:785–801 DOI 10.1007/s00018-010-0566-5 Cellular and Molecular Life Sciences REVIEW The plasmin–antiplasmin system: structural and functional aspects Johann Schaller • Simon S. Gerber Received: 13 April 2010 / Revised: 3 September 2010 / Accepted: 12 October 2010 / Published online: 7 December 2010 Ó Springer Basel AG 2010 Abstract The plasmin–antiplasmin system plays a key Plasminogen activator inhibitors Á a2-Macroglobulin Á role in blood coagulation and fibrinolysis. Plasmin and Multidomain serine proteases a2-antiplasmin are primarily responsible for a controlled and regulated dissolution of the fibrin polymers into solu- Abbreviations ble fragments. However, besides plasmin(ogen) and A2PI a2-Antiplasmin, a2-Plasmin inhibitor a2-antiplasmin the system contains a series of specific CHO Carbohydrate activators and inhibitors. The main physiological activators EGF-like Epidermal growth factor-like of plasminogen are tissue-type plasminogen activator, FN1 Fibronectin type I which is mainly involved in the dissolution of the fibrin K Kringle polymers by plasmin, and urokinase-type plasminogen LBS Lysine binding site activator, which is primarily responsible for the generation LMW Low molecular weight of plasmin activity in the intercellular space. Both activa- a2M a2-Macroglobulin tors are multidomain serine proteases. Besides the main NTP N-terminal peptide of Pgn physiological inhibitor a2-antiplasmin, the plasmin–anti- PAI-1, -2 Plasminogen activator inhibitor 1, 2 plasmin system is also regulated by the general protease Pgn Plasminogen inhibitor a2-macroglobulin, a member of the protease Plm Plasmin inhibitor I39 family.
    [Show full text]
  • Development and Binding Characteristics of Phosphonate Inhibitors of Spla Protease from Staphylococcus Aureus
    Development and binding characteristics of phosphonate inhibitors of SplA protease from Staphylococcus aureus Ewa Burchacka,1 Michal Zdzalik,2 Justyna-Stec Niemczyk,3 Katarzyna Pustelny,3 Grzegorz Popowicz,4 Benedykt Wladyka,3,5 Adam Dubin,3 Jan Potempa,2 Marcin Sienczyk,1 Grzegorz Dubin,2,5* and Jozef Oleksyszyn1 1Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland 2Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland 3Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland 4Max-Planck Institute for Biochemistry, NMR Group, Martinsried, Germany 5Malopolska Centre of Biotechnology, Krakow, Poland Received 24 September 2013; Revised 27 November 2013; Accepted 2 December 2013 DOI: 10.1002/pro.2403 Published online 4 December 2013 proteinscience.org Abstract: Staphylococcus aureus is responsible for a variety of human infections, including life- threatening, systemic conditions. Secreted proteome, including a range of proteases, constitutes the major virulence factor of the bacterium. However, the functions of individual enzymes, in par- ticular SplA protease, remain poorly characterized. Here, we report development of specific inhibi- tors of SplA protease. The design, synthesis, and activity of a series of a-aminoalkylphosphonate diaryl esters and their peptidyl derivatives are described. Potent inhibitors of SplA are reported, which may facilitate future investigation of physiological function of the protease. The binding P P modes of the high-affinity compounds Cbz-Phe -(OC6H424-SO2CH3)2 and Suc-Val-Pro-Phe - (OC6H5)2 are revealed by high-resolution crystal structures of complexes with the protease. Sur- prisingly, the binding mode of both compounds deviates from previously characterized canonical interaction of a-aminoalkylphosphonate peptidyl derivatives and family S1 serine proteases.
    [Show full text]
  • Association Between Production of Fibrinolysin Exudative Epidermitis in Pigs
    Acta vet. scand. 1997, 38, 295-297. Brief Communication Association Between Production of Fibrinolysin and Virulence of Staphylococcus hyicus in Relation to Exudative Epidermitis in Pigs Staphylococcus hyicus is the causative agent of of these is fibrinolysin or staphylokinase. exudative epidermitis (EE) in pigs, character­ Staphylokinase is a protein produced by many ized by a generalized infection of the skin with S. aureus and S. hyicus strains (Devriese et al. greasy exudation and exfoliation (L'Ecuyer 1978, Devriese & Kerckhove 1980) that binds 1966). S. hyicus is a natural part ofthe skin flora to plasminogen and converts it into plasmin that of healthy pigs worldwide (Wegener 1992), and dissolves proteins and fibrinogen. The potential several different strains may simultaneously rple of fibrinolysin in pathogenesis of bacterial colonize the same pig (Wegener 1993a). Both infections is not known, but it has been specu­ virulent and avirulent strains can be present si­ lated that it might help the bacteria in obtaining multaneously on diseased piglets (Wegener et amino acids and in colonization by dissolving al. 1993), and virulent strains can be isolated fibrinogen and other proteins. Production offib ­ from healthy carriers (Devriese 1977, Park & rinolysin has previously been described in S. Kang 1988). The pathogenesis of EE has only hyicus from pigs in different countries in Eu­ been studied in a limited number of studies, but rope (Devriese et al. 1978). This study de­ EE most likely occurs as a consequence of skin scribes the occurrence of fibrinolysin produc­ trauma that exposes the dermis and facilitates tion among virulent and avirulent isolates of S.
    [Show full text]
  • Staphylococcal Manipulation of Host Immune Responses
    REVIEWS Staphylococcal manipulation of host immune responses Vilasack Thammavongsa1,2, Hwan Keun Kim1, Dominique Missiakas1 and Olaf Schneewind1 Abstract | Staphylococcus aureus, a bacterial commensal of the human nares and skin, is a frequent cause of soft tissue and bloodstream infections. A hallmark of staphylococcal infections is their frequent recurrence, even when treated with antibiotics and surgical intervention, which demonstrates the bacterium’s ability to manipulate innate and adaptive immune responses. In this Review, we highlight how S. aureus virulence factors inhibit complement activation, block and destroy phagocytic cells and modify host B cell and T cell responses, and we discuss how these insights might be useful for the development of novel therapies against infections with antibiotic resistant strains such as methicillin-resistant S. aureus. 4 Abscesses Approximately 30% of the human population is contin- signals (that is, chemoattractants and cytokines ). The pathological product of uously colonized with Staphylococcus aureus, whereas Staphylococcal products are detected by immune cells Staphylococcus aureus some individuals are hosts for intermittent colonization1. via Toll-like receptors (TLRs) and G protein-coupled infection: the harbouring of S. aureus typically resides in the nares but is also found receptors, whereas cytokines activate cognate immune a staphylococcal abscess on the skin and in the gastrointestinal tract. Although receptors. Neutrophils answer this call, extravasate from community within a pseudocapsule of fibrin colonization is not a prerequisite for staphylococcal blood vessels, and migrate towards the site of infection deposits that is surrounded by disease, colonized individuals more frequently acquire to phagocytose and kill bacteria or to immobilize and layers of infiltrating immune infections1.
    [Show full text]
  • The Role of Fibrinolytic Regulators in Vascular Dysfunction of Systemic Sclerosis
    International Journal of Molecular Sciences Review The Role of Fibrinolytic Regulators in Vascular Dysfunction of Systemic Sclerosis Yosuke Kanno Department of Clinical Pathological Biochemistry, Faculty of Pharmaceutical Science, Doshisha Women’s College of Liberal Arts, 97-1 Kodo Kyo-tanabe, Kyoto 610-0395, Japan; [email protected]; Tel.: +81-0774-65-8629 Received: 19 November 2018; Accepted: 29 January 2019; Published: 31 January 2019 Abstract: Systemic sclerosis (SSc) is a connective tissue disease of autoimmune origin characterized by vascular dysfunction and extensive fibrosis of the skin and visceral organs. Vascular dysfunction is caused by endothelial cell (EC) apoptosis, defective angiogenesis, defective vasculogenesis, endothelial-to-mesenchymal transition (EndoMT), and coagulation abnormalities, and exacerbates the disease. Fibrinolytic regulators, such as plasminogen (Plg), plasmin, α2-antiplasmin (α2AP), tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA) and its receptor (uPAR), plasminogen activator inhibitor 1 (PAI-1), and angiostatin, are considered to play an important role in the maintenance of endothelial homeostasis, and are associated with the endothelial dysfunction of SSc. This review considers the roles of fibrinolytic factors in vascular dysfunction of SSc. Keywords: Fibrinolytic regulators; SSc; vascular dysfunction 1. Introduction Systemic sclerosis (SSc) is an autoimmune rheumatic disease of unknown etiology that is characterized by vascular dysfunction and fibrosis of the skin and visceral organs as well as peripheral circulatory disturbance [1]. This process usually occurs over many months and years and can lead to organ dysfunction or death. In SSc, vascular disorders are observed from early onset to the appearance of late complications and affect various organs, including the lungs, kidneys, heart, and digital arteries, and exacerbate the disease [2].
    [Show full text]
  • High Level Interferon Alpha-2B Production in Trichoderma Reesei
    Landowski et al. Microb Cell Fact (2016) 15:104 DOI 10.1186/s12934-016-0508-5 Microbial Cell Factories RESEARCH Open Access Enabling low cost biopharmaceuticals: high level interferon alpha‑2b production in Trichoderma reesei Christopher P. Landowski1*, Eero Mustalahti1, Ramon Wahl2, Laurence Croute2, Dhinakaran Sivasiddarthan1, Ann Westerholm‑Parvinen1, Benjamin Sommer2, Christian Ostermeier2, Bernhard Helk2, Juhani Saarinen3 and Markku Saloheimo1 Abstract Background: The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have con‑ structed a series of interferon alpha-2b (IFNα-2b) production strains with 9 protease deletions to gain knowledge for further strain development. Results: We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNα-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein. Conclusions: High levels of IFNα-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving car‑ rier protein cleavage.
    [Show full text]
  • Inhibition of the ATP Synthase Eliminates the Intrinsic Resistance
    Downloaded from RESEARCH ARTICLE crossm mbio.asm.org Inhibition of the ATP Synthase on December 19, 2017 - Published by Eliminates the Intrinsic Resistance of Staphylococcus aureus towards Polymyxins Martin Vestergaard,a Katrine Nøhr-Meldgaard,a Martin Saxtorph Bojer,a Christina Krogsgård Nielsen,b Rikke Louise Meyer,c Christoph Slavetinsky,d Andreas Peschel,d Hanne Ingmera Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, mbio.asm.org Frederiksberg C, Denmarka; Interdisciplinary Nanoscience Center, Aarhus University, Aarhus C, Denmarkb; Department of Bioscience, Aarhus University, Aarhus C, Denmarkc; Interfaculty Institute of Microbiology and Infection Medicine, Infection Biology Section, University of Tübingen, and German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germanyd ABSTRACT Staphylococcus aureus is intrinsically resistant to polymyxins (polymyxin B and colistin), an important class of cationic antimicrobial peptides used in treat- Received 25 June 2017 Accepted 7 August 2017 Published 5 September 2017 ment of Gram-negative bacterial infections. To understand the mechanisms underly- Citation Vestergaard M, Nøhr-Meldgaard K, ing intrinsic polymyxin resistance in S. aureus, we screened the Nebraska Transposon Bojer MS, Krogsgård Nielsen C, Meyer RL, Mutant Library established in S. aureus strain JE2 for increased susceptibility to poly- Slavetinsky C, Peschel A, Ingmer H. 2017. Inhibition of the ATP synthase eliminates the myxin B. Nineteen mutants displayed at least 2-fold reductions in MIC, while the intrinsic resistance of Staphylococcus aureus greatest reductions (8-fold) were observed for mutants with inactivation of either towards polymyxins. mBio 8:e01114-17. graS, graR, vraF,orvraG or the subunits of the ATP synthase (atpA, atpB, atpG,or https://doi.org/10.1128/mBio.01114-17.
    [Show full text]
  • Structural Studies of the Staphylococcus Aureus Ribosome Iskander Khusainov
    Structural studies of the Staphylococcus aureus ribosome Iskander Khusainov To cite this version: Iskander Khusainov. Structural studies of the Staphylococcus aureus ribosome. Biophysics. Université de Strasbourg, 2015. English. NNT : 2015STRAJ071. tel-01403855 HAL Id: tel-01403855 https://tel.archives-ouvertes.fr/tel-01403855 Submitted on 28 Nov 2016 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. UNIVERSITÉ DE STRASBOURG ÉCOLE DOCTORALE des Sciences de la Vie et de la Santé IGBMC - CNRS UMR 7104 - Inserm U 964 THÈSE présentée par : Iskander KHUSAINOV soutenue le : 27 Novembre 2015 pour obtenir le grade de : !"#$%&'($')R%+,-$&.,# '($'0#&1.2!%&3 Discipline/ Spécialité : Biophysique et biologie structurale (Biophysics and Structural biology) Structural studies of the Staphylococcus aureus ribosome Etudes structurales du ribosome de Staphylococcus aureus THÈSE dirigée par : M. YUSUPOV Marat Directeur de recherche, Université de Strasbourg Mme. YUSUPOVA Gulnara Directeur de recherche, Université de Strasbourg RAPPORTEURS : M. JENNER Lasse B. Associate
    [Show full text]