DOCSLIB.ORG

Gelatinase-Mediated Migration and Invasion of Cancer Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Gelatinase-Mediated Migration and Invasion of Cancer Cells View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Helsingin yliopiston digitaalinen arkisto Gelatinase-mediated Migration and Invasion of Cancer Cells Mikael Björklund Department of Biological and Environmental Sciences, Faculty of Biosciences, and Viikki Graduate School in Biosciences, University of Helsinki Academic Dissertation To be presented for public criticism, with the permission of the Faculty of Biosciences, University of Helsinki, in the Lecture Hall 2402 at Viikki Biocenter Bldg. 3, Helsinki, on October 22nd, 2004, at 12 p.m. Helsinki 2004 Supervised by: Docent Erkki Koivunen Department of Biological and Environmental Sciences University of Helsinki Helsinki, Finland Reviewed by: Professor Jorma Keski-Oja. Departments of Pathology and Virology Biomedicum and Haartman Institute University of Helsinki Helsinki, Finland and Professor Jyrki Heino Department of Biochemistry and Food Chemistry University of Turku Turku, Finland Opponent: Professor Antti Vaheri Department of Virology Haartman Institute University of Helsinki Helsinki, Finland ISSN: 1239-9469 ISBN: 952-10-2024-5 (printed) 952-10-2025-3 (PDF) Yliopistopaino Helsinki 2004 2 CONTENTS CONTENTS................................................................................................................................................. 3 LIST OF ORIGINAL PUBLICATIONS .................................................................................................... 5 ABBREVIATIONS...................................................................................................................................... 6 ABSTRACT................................................................................................................................................. 7 INTRODUCTION ....................................................................................................................................... 8 PHAGE DISPLAY ...................................................................................................................................... 8 PHAGE DISPLAY FORMATS.......................................................................................................................... 8 BIOPANNING WITH PHAGE DISPLAY LIBRARIES ............................................................................................ 9 APPLICATIONS OF PEPTIDE DISPLAY.......................................................................................................... 10 ANALYSIS OF PHAGE PEPTIDES ................................................................................................................. 11 Recombinant protein expression using self-splicing inteins.................................................................. 12 BIOSYNTHETIC METHODS FOR INCORPORATION OF NOVEL AMINO ACIDS .................................................... 12 Residue-specific incorporation of non-canonical amino acids by misaminoacylation ........................... 13 Site-directed incorporation of unnatural amino acids.......................................................................... 14 CURRENT APPROACHES TO INCREASE THE CHEMICAL DIVERSITY OF BIOLOGICAL DISPLAY LIBRARIES.......... 15 GELATINASES AND OTHER MATRIX METALLOPROTEINASES................................................. 17 PHYSIOLOGICAL AND PATHOLOGICAL ROLES OF GELATINASES .................................................................. 17 STRUCTURAL FEATURES OF MATRIX METALLOPROTEINASES...................................................................... 18 GELATINASE SUBSTRATES........................................................................................................................ 21 REGULATION OF MMP ACTIVITY ............................................................................................................. 23 Expression and secretion of gelatinases and other MMPs.................................................................... 24 Proenzyme activation ......................................................................................................................... 25 GELATINASE INHIBITORS................................................................................................................... 28 NATURALLY OCCURING GELATINASE INHIBITORS...................................................................................... 28 SYNTHETIC GELATINASE INHIBITORS ........................................................................................................ 31 CELL MIGRATION AND INVASION.................................................................................................... 33 REGULATION OF CELL MIGRATION............................................................................................................ 33 MULTIPLE ROLES OF PROTEINASES IN CELL MIGRATION AND INVASION ...................................................... 35 CELL SURFACE ASSOCIATIONS OF THE GELATINASES ................................................................................. 37 OTHER PROTEASES IN CELL MIGRATION AND INVASION ............................................................................. 40 GELATINASES IN TUMOR PROGRESSION ....................................................................................... 43 ANGIOGENESIS ........................................................................................................................................ 43 METASTASIS............................................................................................................................................ 44 CANCER-ASSOCIATED INFLAMMATION...................................................................................................... 46 DUAL ROLE OF GELATINASES IN CANCER .................................................................................................. 47 THERAPEUTIC POSSIBILITIES WITH THE MMP INHIBITORS .................................................... 48 AIMS OF THIS STUDY............................................................................................................................ 50 MATERIAL AND METHODS................................................................................................................. 51 RESULTS AND DISCUSSION................................................................................................................. 55 BIOSYNTHESIS OF PHAGE DISPLAY PEPTIDES (I, III-IV).............................................................................. 55 INCREASE OF SERUM STABILITY OF GELATINASE INHIBITOR PEPTIDE CTT BY INCORPORATION OF A 5- FLUOROTRYPTOPHAN (I) .......................................................................................................................... 55 INCORPORATION OF AMINO ACID ANALOGUES INTO PHAGE DISPLAY LIBRARIES (II) .................................... 56 3 INHIBITION OF MMP-9 INTERACTION WITH αM INTEGRIN I DOMAIN BY PHAGE DISPLAY PEPTIDES CONTAING A DDGW MOTIF (III) ...............................................................................................................................56 IDENTIFICATION OF DOMAIN-SPECIFIC LIGANDS OF MMP-9 THAT INHIBIT TUMOR CELL MIGRATION AND INVASION (IV) .........................................................................................................................................57 MMP-9 INTERACTS WITH THE UROKINASE-PLASMINOGEN ACTIVATOR RECEPTOR AND THE INTEGRIN β CHAIN (IV).........................................................................................................................................................58 IDENTIFICATION OF A SMALL MOLECULE INHIBITOR OF αMβ2 INTEGRIN-DEPENDENT LEUKEMIA CELL MIGRATION (V)........................................................................................................................................59 CONCLUDING REMARKS.....................................................................................................................61 ACKNOWLEDGEMENTS.......................................................................................................................64 REFERENCES ..........................................................................................................................................65 4 LIST OF ORIGINAL PUBLICATIONS This thesis is based on the following original articles, which are referred to by their Roman numerals in the text. I Björklund, M., Valtanen, H., Savilahti, H. and Koivunen, E: Use of intein-directed peptide biosynthesis to improve serum stability and bioactivity of a gelatinase inhibitory peptide. Combinatorial Chemistry and High Throughput Screening 6, 29-35, 2003. II Björklund, M. and Koivunen, E: Steps towards phage display libraries with an extended amino acid repertoire. Letters in Drug Design & Discovery 1, 163-167, 2004. III Stefanidakis, M., Björklund, M., Ihanus, E., Gahmberg, C. G. and Koivunen, E: Identification of a negatively charged peptide motif within the catalytic domain of progelatinases that mediates binding to leukocyte β2 integrins. Journal of Biological Chemistry, 278, 34674-84, 2003. IV Björklund, M., Heikkilä, P. and Koivunen, E: Peptide inhibition of catalytic and noncatalytic activities of matrix metalloproteinase-9 blocks tumor cell migration and invasion. Journal of Biological Chemistry, 279, 29589-97, 2004. V Björklund, M. and Koivunen, E: A small-molecule stabilizing the active conformation of the αM integrin I domain inhibits leukemia cell migration. Submitted, 2004. The original articles
Load more

Recommended publications
  • Proteases, Mucus, and Mucosal Immunity in Chronic Lung Disease
    International Journal of Molecular Sciences Review Proteases, Mucus, and Mucosal Immunity in Chronic Lung Disease Michael C. McKelvey 1 , Ryan Brown 1, Sinéad Ryan 1, Marcus A. Mall 2,3,4 , Sinéad Weldon 1 and Clifford C. Taggart 1,* 1 Airway Innate Immunity Research (AiiR) Group, Wellcome-Wolfson Institute for Experimental Medicine, Queen’s University Belfast, Belfast BT9 7BL, UK; [email protected] (M.C.M.); [email protected] (R.B.); [email protected] (S.R.); [email protected] (S.W.) 2 Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Charité—Universitätsmedizin Berlin, 13353 Berlin, Germany; [email protected] 3 Berlin Institute of Health (BIH), 10178 Berlin, Germany 4 German Center for Lung Research (DZL), 35392 Gießen, Germany * Correspondence: [email protected]; Tel.: +44-289097-6383 Abstract: Dysregulated protease activity has long been implicated in the pathogenesis of chronic lung diseases and especially in conditions that display mucus obstruction, such as chronic obstructive pulmonary disease, cystic fibrosis, and non-cystic fibrosis bronchiectasis. However, our appreciation of the roles of proteases in various aspects of such diseases continues to grow. Patients with muco- obstructive lung disease experience progressive spirals of inflammation, mucostasis, airway infection and lung function decline. Some therapies exist for the treatment of these symptoms, but they are unable to halt disease progression and patients may benefit from novel adjunct therapies. In this review, we highlight how proteases act as multifunctional enzymes that are vital for normal Citation: McKelvey, M.C.; Brown, R.; airway homeostasis but, when their activity becomes immoderate, also directly contribute to airway Ryan, S.; Mall, M.A.; Weldon, S.; dysfunction, and impair the processes that could resolve disease.
    [Show full text]
  • Metalloproteinase-9 and Chemotaxis Inflammatory Cell Production of Matrix AQARSAASKVKVSMKF, Induces 5, Α a Site on Laminin
    A Site on Laminin α5, AQARSAASKVKVSMKF, Induces Inflammatory Cell Production of Matrix Metalloproteinase-9 and Chemotaxis This information is current as of September 25, 2021. Tracy L. Adair-Kirk, Jeffrey J. Atkinson, Thomas J. Broekelmann, Masayuki Doi, Karl Tryggvason, Jeffrey H. Miner, Robert P. Mecham and Robert M. Senior J Immunol 2003; 171:398-406; ; doi: 10.4049/jimmunol.171.1.398 Downloaded from http://www.jimmunol.org/content/171/1/398 References This article cites 64 articles, 24 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/171/1/398.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 25, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2003 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology A Site on Laminin ␣5, AQARSAASKVKVSMKF, Induces Inflammatory Cell Production of Matrix Metalloproteinase-9 and Chemotaxis1 Tracy L. Adair-Kirk,* Jeffrey J. Atkinson,* Thomas J.
    [Show full text]
  • What Are the Roles of Metalloproteinases in Cartilage and Bone Damage? G Murphy, M H Lee
    iv44 Ann Rheum Dis: first published as 10.1136/ard.2005.042465 on 20 October 2005. Downloaded from REPORT What are the roles of metalloproteinases in cartilage and bone damage? G Murphy, M H Lee ............................................................................................................................... Ann Rheum Dis 2005;64:iv44–iv47. doi: 10.1136/ard.2005.042465 enzyme moiety into an upper and a lower subdomain. A A role for metalloproteinases in the pathological destruction common five stranded beta-sheet and two alpha-helices are in diseases such as rheumatoid arthritis and osteoarthritis, always found in the upper subdomain with a further C- and the irreversible nature of the ensuing cartilage and bone terminal helix in the lower subdomain. The catalytic sites of damage, have been the focus of much investigation for the metalloproteinases, especially the MMPs, have been several decades. This has led to the development of broad targeted for the development of low molecular weight spectrum metalloproteinase inhibitors as potential therapeu- synthetic inhibitors with a zinc chelating moiety. Inhibitors tics. More recently it has been appreciated that several able to fully differentiate between individual enzymes have families of zinc dependent proteinases play significant and not been identified thus far, although a reasonable level of varied roles in the biology of the resident cells in these tissues, discrimination is now being achieved in some cases.7 Each orchestrating development, remodelling, and subsequent family does, however, have other unique domains with pathological processes. They also play key roles in the numerous roles, including the determination of physiological activity of inflammatory cells. The task of elucidating the substrate specificity, ECM, or cell surface localisation (fig 1).
    [Show full text]
  • High Serum Levels of Matrix Metalloproteinase-9 and Matrix
    Imaging, Diagnosis, Prognosis High Serum Levels of Matrix Metalloproteinase-9 and Matrix Metalloproteinase-1Are Associated with Rapid Progression in Patients with Metastatic Melanoma Johanna Nikkola,1, 3 PiaVihinen,1, 3 Meri-SiskoVuoristo,4 Pirkko Kellokumpu-Lehtinen,4 Veli-Matti Ka« ha« ri,2 and Seppo Pyrho« nen1, 3 Abstract Purpose: Matrixmetalloproteinases (MMP) are proteolytic enzymes that play an important role in various aspects of cancer progression. In the present work, we have studied the prognostic significance of serum levels of gelatinase B (MMP-9), collagenase-1 (MMP-1), and collagenase- 3 (MMP-13) in patients with advanced melanoma. Experimental Design:Total pretreatment serum levels of MMP-9 in 71patients and MMP-1and MMP-13 in 48 patients were determined by an assay system based on ELISA. Total MMP levels were also assessed in eight healthy controls. The active and latent forms of MMPs were defined by usingWestern blot analysis and gelatin zymography. Results: Patients with high serum levels of MMP-9 (z376.6 ng/mL; n = 19) had significantly poorer overall survival (OS) than patients with lower serum MMP-9 levels (n =52;medianOS, 29.1versus 45.2 months; P = 0.033). High MMP-9 levels were also associated with visceral or bone metastasis (P = 0.027), elevated serum alkaline phosphatase level (P = 0.0009), and presence of liver metastases (P =0.032).SerumlevelsofMMP-1andMMP-13didnotcorrelate with OS. MMP-1and MMP-9 were found mainly in latent forms in serum, whereas the majority of MMP-13 in serum was active (48 kDa) form. MMP-13 was found more often in active form in patients (mean, 99% of the total MMP-13 level) than in controls (mean, 84% of the total MMP-13 level; P < 0.0001).
    [Show full text]
  • Cloning of a Salivary Gland Metalloprotease And
    University of Rhode Island DigitalCommons@URI Past Departments Faculty Publications (CELS) College of the Environment and Life Sciences 2003 Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme disease tick vector Ixodes scapularis Ivo M.B. Francischetti Thomas N. Mather University of Rhode Island, [email protected] José M.C. Ribeiro Follow this and additional works at: https://digitalcommons.uri.edu/cels_past_depts_facpubs Citation/Publisher Attribution Francischetti, I. M.B., Mather, T. N., & Ribeiro, J. M.C. (2003). Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme disease tick vector Ixodes scapularis. Biochemical and Biophysical Research Communications, 305(4), 869-875. doi: 10.1016/S0006-291X(03)00857-X Available at: https://doi.org/10.1016/S0006-291X(03)00857-X This Article is brought to you for free and open access by the College of the Environment and Life Sciences at DigitalCommons@URI. It has been accepted for inclusion in Past Departments Faculty Publications (CELS) by an authorized administrator of DigitalCommons@URI. For more information, please contact [email protected]. NIH Public Access Author Manuscript Biochem Biophys Res Commun. Author manuscript; available in PMC 2010 July 14. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Biochem Biophys Res Commun. 2003 June 13; 305(4): 869±875. Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme Disease tick vector Ixodes scapularis Ivo M.
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • Gent Forms of Metalloproteinases in Hydra
    Cell Research (2002); 12(3-4):163-176 http://www.cell-research.com REVIEW Structure, expression, and developmental function of early diver- gent forms of metalloproteinases in Hydra 1 2 3 4 MICHAEL P SARRAS JR , LI YAN , ALEXEY LEONTOVICH , JIN SONG ZHANG 1 Department of Anatomy and Cell Biology University of Kansas Medical Center Kansas City, Kansas 66160- 7400, USA 2 Centocor, Malvern, PA 19355, USA 3 Department of Experimental Pathology, Mayo Clinic, Rochester, MN 55904, USA 4 Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66047, USA ABSTRACT Metalloproteinases have a critical role in a broad spectrum of cellular processes ranging from the breakdown of extracellular matrix to the processing of signal transduction-related proteins. These hydro- lytic functions underlie a variety of mechanisms related to developmental processes as well as disease states. Structural analysis of metalloproteinases from both invertebrate and vertebrate species indicates that these enzymes are highly conserved and arose early during metazoan evolution. In this regard, studies from various laboratories have reported that a number of classes of metalloproteinases are found in hydra, a member of Cnidaria, the second oldest of existing animal phyla. These studies demonstrate that the hydra genome contains at least three classes of metalloproteinases to include members of the 1) astacin class, 2) matrix metalloproteinase class, and 3) neprilysin class. Functional studies indicate that these metalloproteinases play diverse and important roles in hydra morphogenesis and cell differentiation as well as specialized functions in adult polyps. This article will review the structure, expression, and function of these metalloproteinases in hydra. Key words: Hydra, metalloproteinases, development, astacin, matrix metalloproteinases, endothelin.
    [Show full text]
  • 9/Gelatinase B Reduce NK Cell-Mediated Cytotoxicity
    in vivo 22 : 593-598 (2008) A High Concentration of MMP-2/ Gelatinase A and MMP- 9/ Gelatinase B Reduce NK Cell-mediated Cytotoxicity against an Oral Squamous Cell Carcinoma Cell Line BU-KYU LEE 1, MI-JUNG KIM 2, HA-SOON JANG 1, HEE-RAN LEE 2, KANG-MIN AHN 1, JONG-HO LEE 3, PILL-HOON CHOUNG 3 and MYUNG-JIN KIM 3 Departments of 1Oral and Maxillofacial Surgery and 2Cell Biology, Asan Institute for Life Sciences, Asan Medical Center, College of Medicine, Ulsan University, Songpa-ku, 138-040, Seoul; 3Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University, Jongno-ku, 110-768, Seoul, Korea Abstract. Background: Recent studies have shown that advanced state of the disease had generally reduced host matrix metalloproteinases (MMPs) from tumors influence the immune function (3, 4). Nevertheless, the exact role of host immune system to reduce antitumor activity. The aim of immune cells, and of the immune system in general, in the this study was to examine the influence of MMP-2 and development of OSCC and in tumor progression remains MMP-9 on the natural killer (NK) cell. Materials and ambiguous. While the initiation of the process of Methods: NK cells were pretreated with either MMP-2 or tumorigenesis is clearly linked to carcinogens ( i.e. tobacco MMP-9 in the experimental group but not in the control or alcohol), its progression through a series of discrete group. NK cell cytotoxicity against oral squamous cell genetic changes results in the emergence of a tumor that is carcinoma cells (OSCC) were examined using the [Cr 51 ] resistant to immune effector cells (5, 6).
    [Show full text]
  • Signal Transduction, Quorum-Sensing, and Extracellular Protease Activity in Enterococcus Faecalis Biofilm Formation
    Signal Transduction, Quorum-Sensing, and Extracellular Protease Activity in Enterococcus faecalis Biofilm Formation The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Carniol, K., and M. S. Gilmore. 2004. Signal Transduction, Quorum- Sensing, and Extracellular Protease Activity in Enterococcus Faecalis Biofilm Formation. Journal of Bacteriology 186, no. 24: 8161–8163. doi:10.1128/jb.186.24.8161-8163.2004. Published Version doi:10.1128/JB.186.24.8161-8163.2004 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:33867369 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA JOURNAL OF BACTERIOLOGY, Dec. 2004, p. 8161–8163 Vol. 186, No. 24 0021-9193/04/$08.00ϩ0 DOI: 10.1128/JB.186.24.8161–8163.2004 Copyright © 2004, American Society for Microbiology. All Rights Reserved. GUEST COMMENTARY Signal Transduction, Quorum-Sensing, and Extracellular Protease Activity in Enterococcus faecalis Biofilm Formation Karen Carniol1,2 and Michael S. Gilmore1,2* Department of Ophthalmology, Harvard Medical School,1 and The Schepens Eye Research Institute,2 Boston, Massachusetts Biofilms are surface-attached communities of bacteria, en- sponse regulator proteins (10). Only one of the mutants gen- cased in an extracellular matrix of secreted proteins, carbohy- erated, fsrA, impaired the ability of E. faecalis strain V583A to drates, and/or DNA, that assume phenotypes distinct from form biofilms in vitro.
    [Show full text]
  • An Adverse Role for Matrix Metalloproteinase 12 After Spinal Cord Injury in Mice
    The Journal of Neuroscience, November 5, 2003 • 23(31):10107–10115 • 10107 Development/Plasticity/Repair An Adverse Role for Matrix Metalloproteinase 12 after Spinal Cord Injury in Mice Jennifer E. A. Wells,1 Tiffany K. Rice,1 Robert K. Nuttall,3 Dylan R. Edwards,3 Hakima Zekki,4 Serge Rivest,4 V. Wee Yong1,2 Departments of 1Clinical Neurosciences and 2Oncology, Faculty of Medicine, University of Calgary, Calgary, Alberta T2N 4N1, Canada, 3School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, United Kingdom, and 4Laboratory of Molecular Endocrinology and Department of Anatomy and Physiology, Laval University, Quebec, Quebec G1V 4G2, Canada We investigated the role of matrix metalloproteinases (MMPs) in acute spinal cord injury (SCI). Transcripts encoding 22 of the 23 known mammalian MMPs were measured in the mouse spinal cord at various time points after injury. Although there were significant changes in the expression levels of multiple MMPs, MMP-12 was increased 189-fold over normal levels, the highest of all MMPs examined. To evaluate the role of MMP-12 in SCI, spinal cord compression was performed in wild-type (WT) and MMP-12 null mice. Behavioral analyses were conducted for 4 weeks using the Basso–Beattie–Bresnahan (BBB) locomotor rating scale as well as the inclined plane test. The results show that MMP-12 null mice exhibited significantly improved functional recovery compared with WT controls. Twenty-eight days after injury, the BBB score in the MMP-12 group was 7, representing extensive movement of all three hindlimb joints, compared with 4 in the WT group, representing only slight movement of these joints.
    [Show full text]
  • (Matrix Metalloprotease-9) Activity Reduces Cellular Inflammation and Restores Function of Transplanted Pancreatic Islets
    ORIGINAL ARTICLE Inhibition of Gelatinase B (Matrix Metalloprotease-9) Activity Reduces Cellular Inflammation and Restores Function of Transplanted Pancreatic Islets Neelam Lingwal,1 Manju Padmasekar,1 Balaji Samikannu,1 Reinhard G. Bretzel,1 Klaus T. Preissner,2 and Thomas Linn1 Islet transplantation provides an approach to compensate for loss proteolytic activation occurs in the pericellular and extra- of insulin-producing cells in patients with type 1 diabetes. How- cellular space. In two of the secreted MMPs also designated ever, the intraportal route of transplantation is associated with gelatinases, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), instant inflammatory reactions to the graft and subsequent islet the catalytic zinc-carrying domain contains a structural mod- destruction as well. Although matrix metalloprotease (MMP)-2 ule of three fibronectin-like repeats interacting with elastin and -9 are involved in both remodeling of extracellular matrix and and types I, III, and IV gelatins when activated and facilitating leukocyte migration, their influence on the outcome of islet trans- their degradation within connective tissue matrices. plantation has not been characterized. We observed comparable Both neutrophils and macrophages express and secrete MMP-2 mRNA expressions in control and transplanted groups of gelatinases (10,11,13). Previous studies have shown that mice, whereas MMP-9 mRNA and protein expression levels in- fi creased after islet transplantation. Immunostaining for CD11b such cells contribute to the rst line of defense following (Mac-1)-expressing leukocytes (macrophage, neutrophils) and islet transplantation (3,4). Moreover, elevation of MMP-9 Ly6G (neutrophils) revealed substantially reduced inflammatory plasma levels was observed in diabetic patients (14), cell migration into islet-transplanted liver in MMP-9 knockout whereas in mice with acute pancreatitis, trypsin induced recipients.
    [Show full text]
  • Proteolytic Enzymes in Grass Pollen and Their Relationship to Allergenic Proteins
    Proteolytic Enzymes in Grass Pollen and their Relationship to Allergenic Proteins By Rohit G. Saldanha A thesis submitted in fulfilment of the requirements for the degree of Masters by Research Faculty of Medicine The University of New South Wales March 2005 TABLE OF CONTENTS TABLE OF CONTENTS 1 LIST OF FIGURES 6 LIST OF TABLES 8 LIST OF TABLES 8 ABBREVIATIONS 8 ACKNOWLEDGEMENTS 11 PUBLISHED WORK FROM THIS THESIS 12 ABSTRACT 13 1. ASTHMA AND SENSITISATION IN ALLERGIC DISEASES 14 1.1 Defining Asthma and its Clinical Presentation 14 1.2 Inflammatory Responses in Asthma 15 1.2.1 The Early Phase Response 15 1.2.2 The Late Phase Reaction 16 1.3 Effects of Airway Inflammation 16 1.3.1 Respiratory Epithelium 16 1.3.2 Airway Remodelling 17 1.4 Classification of Asthma 18 1.4.1 Extrinsic Asthma 19 1.4.2 Intrinsic Asthma 19 1.5 Prevalence of Asthma 20 1.6 Immunological Sensitisation 22 1.7 Antigen Presentation and development of T cell Responses. 22 1.8 Factors Influencing T cell Activation Responses 25 1.8.1 Co-Stimulatory Interactions 25 1.8.2 Cognate Cellular Interactions 26 1.8.3 Soluble Pro-inflammatory Factors 26 1.9 Intracellular Signalling Mechanisms Regulating T cell Differentiation 30 2 POLLEN ALLERGENS AND THEIR RELATIONSHIP TO PROTEOLYTIC ENZYMES 33 1 2.1 The Role of Pollen Allergens in Asthma 33 2.2 Environmental Factors influencing Pollen Exposure 33 2.3 Classification of Pollen Sources 35 2.3.1 Taxonomy of Pollen Sources 35 2.3.2 Cross-Reactivity between different Pollen Allergens 40 2.4 Classification of Pollen Allergens 41 2.4.1
    [Show full text]