Toxin Instability and Its Role in Toxin Translocation from the Endoplasmic Reticulum to the Cytosol
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Pertussis Toxin
Pertussis Toxin Publication Number MAN0004270 Revision Date 09 May 2011 Catalog Number: PHZ1174 Quantity: 50 μg Lot Number: See product label. Appearance: Lyophilized solid. Origin: Bordetella pertussis. Purity: >99%. This preparation migrates as five distinct bands when analyzed by SDS–Urea PAGE. The five bands correspond to one A protomer subunit, designated S1 (Mr=26.2 kDa), and four B oligomer subunits, designated S2- S5 (Mr’s= 21.9, 21.9, 12.1, and 10.9 kDa, respectively). Summary: Islet-activating protein Pertussis toxin consists of an A protomer subunit (S1) which possesses both NAD+ glycohydrolase and ADP–ribosyltransferase activities, and B oligomer subunits (S2, S3, S4, and S5) which are responsible for attachment of the native toxin to eukaryotic cell surfaces. Pertussis toxin uncouples G proteins from receptors by ADP ribosylating a cysteine residue near the carboxyl terminus of the α subunit. Biological Activity: The lowest concentration which produces a clustered growth pattern with CHO cells is 0.03 ng/mL. The adenylate cyclase activity of this preparation is 20.1 picomoles/minute/μg in the presence of 1 μg calmodulin. Reconstitution Reconstitute the contents of this vial with 500 μL sterile, distilled water. The composition of the solution will be Recommendation: 50 μg Pertussis toxin, 10 mM sodium phosphate, pH 7.0, 50 mM sodium chloride. Because Pertussis toxin is relatively insoluble, the resulting suspension should be made uniform by gentle mixing prior to withdrawing aliquots. It is important to note that this suspension should not be sterile-filtered. This preparation is not activated. For use with intact cells or extracts, activation is not necessary. -
Biological Toxins Fact Sheet
Work with FACT SHEET Biological Toxins The University of Utah Institutional Biosafety Committee (IBC) reviews registrations for work with, possession of, use of, and transfer of acute biological toxins (mammalian LD50 <100 µg/kg body weight) or toxins that fall under the Federal Select Agent Guidelines, as well as the organisms, both natural and recombinant, which produce these toxins Toxins Requiring IBC Registration Laboratory Practices Guidelines for working with biological toxins can be found The following toxins require registration with the IBC. The list in Appendix I of the Biosafety in Microbiological and is not comprehensive. Any toxin with an LD50 greater than 100 µg/kg body weight, or on the select agent list requires Biomedical Laboratories registration. Principal investigators should confirm whether or (http://www.cdc.gov/biosafety/publications/bmbl5/i not the toxins they propose to work with require IBC ndex.htm). These are summarized below. registration by contacting the OEHS Biosafety Officer at [email protected] or 801-581-6590. Routine operations with dilute toxin solutions are Abrin conducted using Biosafety Level 2 (BSL2) practices and Aflatoxin these must be detailed in the IBC protocol and will be Bacillus anthracis edema factor verified during the inspection by OEHS staff prior to IBC Bacillus anthracis lethal toxin Botulinum neurotoxins approval. BSL2 Inspection checklists can be found here Brevetoxin (http://oehs.utah.edu/research-safety/biosafety/ Cholera toxin biosafety-laboratory-audits). All personnel working with Clostridium difficile toxin biological toxins or accessing a toxin laboratory must be Clostridium perfringens toxins Conotoxins trained in the theory and practice of the toxins to be used, Dendrotoxin (DTX) with special emphasis on the nature of the hazards Diacetoxyscirpenol (DAS) associated with laboratory operations and should be Diphtheria toxin familiar with the signs and symptoms of toxin exposure. -
Pertussis Toxin
PLEASE POST THIS PAGE IN AREAS WHERE PERTUSSIS TOXIN IS USED IN RESEARCH LABORATORIES UNIVERSITY OF CALIFORNIA, SAN FRANCISCO ENVIRONMENT, HEALTH AND SAFETY/BIOSAFETY PERTUSSIS TOXIN EXPOSURE/INJURY RESPONSE PROTOCOL Organism or Agent: Pertussis Toxin Exposure Risk; Multiple Endocrine/Metabolic Effects Exposure Hotline Pager: 415/353-7842 (353-STIC) (Available 24 hours) Office of Environment, Health & Safety: 415/476-1300 (Available during work hours) 415/476-1414 or 9-911 (In case of emergency, available 24 hours) EH&S Biosafety Officer 415/514-2824 EH&S Public Health Officer: 415/514-3531 UCSF Occupational Health Services: 415/885-7580 (Available during work hours) California Poison Control: 800/222-1222 SFDPH Emergency Number: 415/554-2830 CDC Emergency Operations: 770/488-7100 _________________________________________________________________________ PROTOCOL SUMMARY In the event of an accidental exposure or injury, the protocol is as follows: 1. Modes of Exposure: a. Skin puncture or injection b. Ingestion c. Contact with mucous membranes (eyes, nose, mouth) d. Contact with non-intact skin e. Exposure to aerosols f. Respiratory exposure from inhalation of toxin 2. First Aid: a. Skin Exposure, immediately go to the sink and thoroughly wash the skin with soap and water. If working with pertussis, decontaminate any exposed skin with an antiseptic scrub solution. b. Skin Wound, immediately go to the sink and thoroughly wash the wound with soap and water and pat dry. c. Splash to Eye(s), Nose or Mouth, immediately flush the area with running water for at least 5- 10 minutes. d. Splash Affecting Garments, remove garments that may have become soiled or contaminated and place them in a double red plastic bag. -
Crystal Structure of a New Heat-Labile Enterotoxin, LT-Iib
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Research Article 665 Crystal structure of a new heat-labile enterotoxin, LT-IIb Focco van den Akker1, Steve Sarfaty1,2, Edda M Twiddy3, Terry D Connell3†, Randall K Holmes3‡ and Wim GJ Hol1,2* Background: Cholera toxin from Vibrio cholerae and the type I heat-labile Addresses: 1Departments of Biological Structure enterotoxins (LT-Is) from Escherichia coli are oligomeric proteins with AB and Biochemistry & Biomolecular Structure 5 Center, University of Washington, 2Howard structures. The type II heat-labile enterotoxins (LT-IIs) from E. coli are structurally Hughes Medical Institute, University of similar to, but antigenically distinct from, the type I enterotoxins. The A subunits Washington, Box 357742, Seattle, WA 98195, of type I and type II enterotoxins are homologous and activate adenylate cyclase USA and 3Department of Microbiology and Immunology, Uniformed Services University of the by ADP-ribosylation of a G protein subunit, Gsa. However, the B subunits of type I and type II enterotoxins differ dramatically in amino acid sequence and Health Sciences, Bethesda, MD 20814, USA. ganglioside-binding specificity. The structure of LT-IIb was determined both as a Present addresses: †Department of Microbiology, prototype for other LT-IIs and to provide additional insights into School of Medicine and Biomedical Sciences, structure/function relationships among members of the heat-labile enterotoxin Buffalo, NY 14214-3078, USA and ‡Department family and the superfamily of ADP-ribosylating protein toxins. of Microbiology, University of Colorado Health Sciences Center, Denver, CO 80262, USA. -
Human Peptides -Defensin-1 and -5 Inhibit Pertussis Toxin
toxins Article Human Peptides α-Defensin-1 and -5 Inhibit Pertussis Toxin Carolin Kling 1, Arto T. Pulliainen 2, Holger Barth 1 and Katharina Ernst 1,* 1 Institute of Pharmacology and Toxicology, Ulm University Medical Center, 89081 Ulm, Germany; [email protected] (C.K.); [email protected] (H.B.) 2 Institute of Biomedicine, Research Unit for Infection and Immunity, University of Turku, FI-20520 Turku, Finland; arto.pulliainen@utu.fi * Correspondence: [email protected] Abstract: Bordetella pertussis causes the severe childhood disease whooping cough, by releasing several toxins, including pertussis toxin (PT) as a major virulence factor. PT is an AB5-type toxin, and consists of the enzymatic A-subunit PTS1 and five B-subunits, which facilitate binding to cells and transport of PTS1 into the cytosol. PTS1 ADP-ribosylates α-subunits of inhibitory G-proteins (Gαi) in the cytosol, which leads to disturbed cAMP signaling. Since PT is crucial for causing severe courses of disease, our aim is to identify new inhibitors against PT, to provide starting points for novel therapeutic approaches. Here, we investigated the effect of human antimicrobial peptides of the defensin family on PT. We demonstrated that PTS1 enzyme activity in vitro was inhibited by α-defensin-1 and -5, but not β-defensin-1. The amount of ADP-ribosylated Gαi was significantly reduced in PT-treated cells, in the presence of α-defensin-1 and -5. Moreover, both α-defensins decreased PT-mediated effects on cAMP signaling in the living cell-based interference in the Gαi- mediated signal transduction (iGIST) assay. -
ADP-Ribosylation with Clostridium Perfringens Iota Toxin
Biochem. J. (1990) 266, 335-339 (Printed in Great Britain) 335 Inhibition of cytochalasin D-stimulated G-actin ATPase by ADP-ribosylation with Clostridium perfringens iota toxin Udo GEIPEL, Ingo JUST and Klaus AKTORIES* Rudolf-Buchheim-Institut fur Pharmakologie der Universitat GieBen, Frankfurterstr. 107, D-6300 GieBen, Federal Republic of Germany Clostridium perfringens iota toxin belongs to a novel family of actin-ADP-ribosylating toxins. The effects of ADP-ribosylation of skeletal muscle actin by Clostridium perfringens iota toxin on cytochalasin D- stimulated actin ATPase activity was studied. Cytochalasin D stimulated actin-catalysed ATP hydrolysis maximally by about 30-fold. ADP-ribosylation of actin completely inhibited cytochalasin D-stimulated ATP hydrolysis. Inhibition of ATPase activity occurred at actin concentrations below the critical concentration (0.1 /iM), at low concentrations of Mg2" (50 ItM) and even in the actin-DNAase I complex, indicating that ADP-ribosylation of actin blocks the ATPase activity of monomeric actin and -that the inhibitory effect is not due to inhibition of the polymerization of actin. INTRODUCTION perfringens type E strain CN5063, which was kindly donated by Dr. S. Thorley (Wellcome Biotech, Various bacterial ADP-ribosylating toxins modify Beckenham, Kent, U.K.) essentially according to the pro- regulatory GTP-binding proteins, thereby affecting cedure described (Stiles & Wilkens, 1986). Cytochalasin eukaryotic cell function. Pertussis toxin and cholera D was obtained from Sigma (Deisenhofen, Germany). toxin ADP-ribosylate GTP-binding proteins involved in DNAase I was a gift from Dr. H. G. Mannherz transmembrane signal transduction (for a review, see (Marburg, Germany). [oc-32P]ATP, [y-32P]ATP and Pfeuffer & Helmreich, 1988). -
Release of Pertussis Toxin and Its Interaction with Outer-Membrane Antigens
Journal of General Microbiology (1987), 133, 2427-2435. Printed in Great Britain 2427 Release of Pertussis Toxin and Its Interaction With Outer-membrane Antigens ByV. Y. PERERA, A. C. WARDLAW AND J. H. FREER* Department of Microbiology, University of Glasgow, Alexander Stone Building, Garscube Estate, Bearsden, Glasgow G61 IQH, UK (Received 16 December 1986 ;revised 22 April 1987) The absence of subunit S3 in cell-associated pertussis toxin (PT) from a mutant of Bordetella pertussis which failed to produce cell-free toxin suggested that this subunit was involved in the release of PT into the culture medium. The addition of methylated P-cyclodextrin (MCD) to the culture medium caused a small but consistent increase in the release of lipopolysaccharide (LPS) by four wild-type strains of B. pertussis. Since previous studies have shown that MCD also enhances the levels of PT in culture supernates, it seemed probable that the increased shedding of outer-membrane vesicles (OMV) may explain the increased levels of both cell-free PT and LPS. Release of PT was inhibited in media buffered with HEPES but was unaffected in Tris/HCl buffer. This suggested that in addition to shedding of the outer membrane, increased permeability and greater destabilization of the outer membrane, as caused by Tris/HCl buffer, may be important in the release of PT. Our data do not support the idea that PT is packaged into OMV because only an insignificant proportion (0.01 %) of the total cell-free PT was associated with LPS. The association of PT with small micelles derived from outer-membrane amphiphiles may be more important since the LPS content of PT purified from culture supernates (containing no large OMV) was nearly 18% by weight. -
Safe Handling of Acutely Toxic Chemicals Safe Handling of Acutely
Safe Handling of Acutely Toxic Chemicals , Mutagens, Teratogens and Reproductive Toxins October 12, 2011 BSBy Sco ttBthlltt Batcheller R&D Manager Milwaukee WI Hazards Classes for Chemicals Flammables • Risk of ignition in air when in contact with common energy sources Corrosives • Generally destructive to materials and tissues Energetic and Reactive Materials • Sudden release of destructive energy possible (e.g. fire, heat, pressure) Toxic Substances • Interaction with cells and organs may lead to tissue damage • EfftEffects are t tilltypically not general ltllti to all tissues, bttbut target tdted to specifi c ones • Examples: – Cancers – Organ diseases – Inflammation, skin rashes – Debilitation from long-term Poison Acute Cancer, health or accumulation with delayed (ingestion) risk reproductive risk emergence 2 Toxic Substances Are All Around Us Pollutants Natural toxins • Cigare tte smo ke • V(kidbt)Venoms (snakes, spiders, bees, etc.) • Automotive exhaust • Poison ivy Common Chemicals • Botulinum toxin • Pesticides • Ricin • Fluorescent lights (mercury) • Radon gas • Asbestos insulation • Arsenic and heavy metals • BPA ((pBisphenol A used in some in ground water plastics) 3 Application at UNL Chemicals in Chemistry Labs Toxin-producing Microorganisms • Chloro form • FiFungi • Formaldehyde • Staphylococcus species • Acetonitrile • Shiga-toxin from E. coli • Benzene Select Agent Toxins (see register) • Sodium azide • Botulinum neurotoxins • Osmium/arsenic/cadmium salts • T-2 toxin Chemicals in Biology Labs • Tetrodotoxin • Phenol -
Instability of Toxin a Subunit of AB5 Toxins in the Bacterial Periplasm Caused by Deficiency of Their Cognate B Subunits
Biochimica et Biophysica Acta 1808 (2011) 2359–2365 Contents lists available at ScienceDirect Biochimica et Biophysica Acta journal homepage: www.elsevier.com/locate/bbamem Instability of toxin A subunit of AB5 toxins in the bacterial periplasm caused by deficiency of their cognate B subunits Sang-Hyun Kim a, Su Hyang Ryu a, Sang-Ho Lee a, Yong-Hoon Lee a, Sang-Rae Lee a, Jae-Won Huh a, Sun-Uk Kim a, Ekyune Kim d, Sunghyun Kim b, Sangyong Jon b, Russell E. Bishop c,⁎, Kyu-Tae Chang a,⁎⁎ a The National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Ochang, Cheongwon, Chungbuk 363–883, Republic of Korea b School of Life Science, Gwangju Institute of Science and Technology (GIST), 1 Oryong-dong, Gwangju 500–712, Republic of Korea c Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada L8N 3Z5 d College of Pharmacy, Catholic University of Daegu, Hayang-eup, Gyeongsan, Gyeongbuk 712-702, Republic of Korea article info abstract Article history: Shiga toxin (STx) belongs to the AB5 toxin family and is transiently localized in the periplasm before secretion Received 26 April 2011 into the extracellular milieu. While producing outer membrane vesicles (OMVs) containing only A subunit of Received in revised form 3 June 2011 the toxin (STxA), we created specific STx1B- and STx2B-deficient mutants of E. coli O157:H7. Surprisingly, Accepted 23 June 2011 STxA subunit was absent in the OMVs and periplasm of the STxB-deficient mutants. In parallel, the A subunit Available online 5 July 2011 of heat-labile toxin (LT) of enterotoxigenic E. -
Impact of Bacterial Toxins in the Lungs
toxins Review Impact of Bacterial Toxins in the Lungs 1,2,3, , 4,5, 3 2 Rudolf Lucas * y, Yalda Hadizamani y, Joyce Gonzales , Boris Gorshkov , Thomas Bodmer 6, Yves Berthiaume 7, Ueli Moehrlen 8, Hartmut Lode 9, Hanno Huwer 10, Martina Hudel 11, Mobarak Abu Mraheil 11, Haroldo Alfredo Flores Toque 1,2, 11 4,5,12,13, , Trinad Chakraborty and Jürg Hamacher * y 1 Pharmacology and Toxicology, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA; hfl[email protected] 2 Vascular Biology Center, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA; [email protected] 3 Department of Medicine and Division of Pulmonary Critical Care Medicine, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA; [email protected] 4 Lungen-und Atmungsstiftung, Bern, 3012 Bern, Switzerland; [email protected] 5 Pneumology, Clinic for General Internal Medicine, Lindenhofspital Bern, 3012 Bern, Switzerland 6 Labormedizinisches Zentrum Dr. Risch, Waldeggstr. 37 CH-3097 Liebefeld, Switzerland; [email protected] 7 Department of Medicine, Faculty of Medicine, Université de Montréal, Montréal, QC H3T 1J4, Canada; [email protected] 8 Pediatric Surgery, University Children’s Hospital, Zürich, Steinwiesstrasse 75, CH-8032 Zürch, Switzerland; [email protected] 9 Insitut für klinische Pharmakologie, Charité, Universitätsklinikum Berlin, Reichsstrasse 2, D-14052 Berlin, Germany; [email protected] 10 Department of Cardiothoracic Surgery, Voelklingen Heart Center, 66333 -
Cellular Activity of Salmonella Typhimurium Artab Toxin and Its Receptor-Binding Subunit
toxins Article Cellular Activity of Salmonella Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit Elise Overgaard 1, Brad Morris 2, Omid Mohammad Mousa 2 , Emily Price 3, Adriana Rodriguez 2, Leyla Cufurovic 2, Richard S. Beard 4 and Juliette K. Tinker 1,2,* 1 Biomolecular Sciences Graduate Program, Boise State University, Boise, ID 83725, USA; [email protected] 2 Department of Biology, Boise State University, Boise, ID 83725, USA; [email protected] (B.M.); [email protected] (O.M.M.); [email protected] (A.R.); [email protected] (L.C.) 3 Idaho Veterans Research and Education Foundation, Infectious Diseases Section, Boise, ID 83702, USA; [email protected] 4 Biomolecular Research Center, Boise State University, Boise, ID 83725, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-208-426-5472 Abstract: Salmonellosis is among the most reported foodborne illnesses in the United States. The Salmonella enterica Typhimurium DT104 phage type, which is associated with multidrug-resistant disease in humans and animals, possesses an ADP-ribosylating toxin called ArtAB. Full-length artAB has been found on a number of broad-host-range non-typhoidal Salmonella species and serovars. ArtAB is also homologous to many AB5 toxins from diverse Gram-negative pathogens, including cholera toxin (CT) and pertussis toxin (PT), and may be involved in Salmonella pathogenesis, however, in vitro cellular toxicity of ArtAB has not been characterized. artAB was cloned into E. coli and initially isolated using a histidine tag (ArtABHIS) and nickel chromatography. ArtABHIS was found to bind to African green monkey kidney epithelial (Vero) cells using confocal microscopy and to Citation: Overgaard, E.; Morris, B.; interact with glycans present on fetuin and monosialotetrahexosylganglioside (GM1) using ELISA. -
Virtuous Aspects of Vicious Bacterial Toxins
Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 2015, 7(6):279-289 ISSN : 0975-7384 Review Article CODEN(USA) : JCPRC5 Virtuous aspects of vicious bacterial toxins Pankaj Gautam, Pranjali Gupta, Promila Sharma and Pranshu Dangwal Department of Biotechnology, Graphic Era University, 566/6, Bell Road, Clement Town, Dehradun, Uttarakhand(India) _____________________________________________________________________________________________ ABSTRACT Pathogenic bacteria exert their harmful effects in host through a cascade of virulence factors: exotoxins, endotoxin, invasions proteins and the like. Various toxins secreted by different bacteria not only subvert the host intracellular signaling pathways but also have unique affinity for specific host cells. Detailed information has been accumulated over the years regarding the purification, chemical characterization, enzymic action and architecture of various bacterial toxins. Toxins ability to bind to the specific target cells makes them good candidate for drug delivery and in cancer therapeutics. A number of immunotoxins; hybrid molecules of bacterial toxins and antibodies, have wide applications in cancer and are currently under clinical trial. Advances in genomics and proteomics have created a wealth of information related to the nucleotide and protein sequences of numerous toxins and different databases (DBETH, VFDB, BETAWRAP, RASTA) are available with elaborate information thereof. In present review we have provided a brief account of therapeutic aspect of bacterial toxins and an effort has been made to facilitate the reader’s understanding as to how we can turn the vicious toxin into virtuous toxin for human benefits. Key words: .Bacterial Toxin, Immunotoxins, AB 5, A2B5, Toxin databases, Therapeutic toxin. _____________________________________________________________________________________________ Bacterial toxins; the soluble antigens, secreted by a number of pathogenic bacteria has a standing reputation of being a poison secreted during the course of pathogenesis.