DOCSLIB.ORG

Mechanistic Studies of Two Phosphatase Enzymes Involved in Inostiol Metabolism

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mechanistic Studies of Two Phosphatase Enzymes Involved in Inostiol Metabolism Mechanistic studies of two phosphatase enzymes involved in inostiol metabolism Author: Yang Wei Persistent link: http://hdl.handle.net/2345/3047 This work is posted on eScholarship@BC, Boston College University Libraries. Boston College Electronic Thesis or Dissertation, 2013 Copyright is held by the author, with all rights reserved, unless otherwise noted. Boston College The Graduate School of Arts and Sciences Department of Chemistry MECHANIC STUDIES OF TWO PHOSPHATASE ENZYMES INVOLVED IN INOSITOL METABOLISM a dissertation by YANG WEI submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy May, 2013 © copyright by YANG WEI 2013 Mechanic studies of two phosphatase enzymes involved in inositol metabolism by Yang Wei Under the direction of Dr. Mary F. Roberts Abstract Inositol-containing molecules and inositol phosphatases have diverse roles in cells. One of the inositol phospholipids phosphatases, PTEN (Phosphatase and Tensin Homolog deleted on Chromosome Ten), is a tumor suppressor and antagonizes the PI3K signaling pathway by dephosphorylating PI(3,4,5)P3 at the 3 position of the inositol ring. In testing predictions of a molecular dynamics simulation, a hydrophobic site adjacent to the active site on PTEN was identified and verified by protein kinetic studies. This hydrophobic site plays an important role in substrate and substrate analogue binding with one of residues, Arg47, critical for PTEN phosphatase activity. Mutations of Arg47 reduced enzyme activities toward both the short-chain substrate as monomers and micelles and long-chain phospholipid presented in vesicles. PI(4,5)P2, the product of PI(3,4,5)P3 dephosphorylation, activates PTEN. Studies by others suggested this occurred when the product was bound to the N-terminal region of the protein (not visible in the crystal structure). However, no direct proof of this existed. The effect of PI(4,5)P2 on PTEN enzyme activities in different substrates systems was studied. 31P NMR was used to probe 31 the spatial location and functional role of PI(4,5)P2 binding site. The fixed field P NMR and high resolution field cycling 31P NMR results indicated there are discrete sites for both substrate and activator lipids on PTEN, and both of sites are spatially separate from the hydrophobic site. The active site, adjacent hydrophobic site, and N-terminal activator binding site worked synergistically to regulate PTEN interacting with the membrane. Thermophilic and hyperthermophilic archaea and bacteria thrive at high temperatures. They often accumulate small organic molecules, called compatible solutes or osmolytes, to protect proteins from thermal denaturation. The thermoprotection mechanism of compatible solutes was explored using inositol monophosphatase (IMPase) from Archaeoglobus fulgidus as the model protein. The protective effect of unusual compatible solutes, di-inositol-1,1’-phosphate (DIP) and diglycerol phosphate (DGP), as well as common compatible solutes glutamate and other anions, on the IMPase thermostability was studied. Specific binding sites of glutamate ions on the IMPase were identified by crystallography and field cycling NMR. However, mutations at these discrete binding sites did not eliminate the thermoprotection, but reduced the thermal stability (Tm) of the protein. Our results indicate the specific binding of osmolytes to the protein exists, but they do not account for the thermoprotection. This dissertation is dedicated to my parents Xiaofei Wei, Lin Yuan and my fiancé Kai Hong i Acknowledgement First, I would like to express my deep gratitude to my advisor, Dr. Mary F. Roberts, for her continuous guidance and support throughout my graduate study. Without her enthusiastic encouragement and confidence, this work would not have been completed. I always appreciate her kindness, patience and generosity. I would like to thank Dr. Goran Krilov and Dr. Qin Wang for doing the simulations of PTEN and discussing this project. I wish to thank Dr. Rebecca Goldstein in the Roberts group for solving the crystal structures of IMPase and collaborations on this project. I wish to thank Dr. Scott Miller, Dr. Yingju Xu and Dr. Christina Longo for providing synthetic deoxy-PI and DIP compounds. I would like to thank Dr. Eranthie Weerapana and Julianne Martell for doing mass spectrometry and data analysis of PTEN spin-labeling samples. I also thank Dr. Weerapana for being my committee member and spending time to read my thesis. I would like to thank Dr. Patrick Wintrode for letting me work in his laboratory to use hydrogen exchange mass spectrometry to study PTEN and PI-PLC. I would like to thank Dr. John Boylan and Dr. Thusitha Jayasundera at Boston College and Dr. Alfred Redfield at Brandeis University for maintaining the NMR facility to make my extensive usage of NMR possible. I wish to thank Dr. Jianmin Gao for letting me use some equipment in his laboratory and being my committee member to read my thesis. ii I wish to thank Dr. Olga Gursky for being a committee member and reading my thesis. I would like to thank Dr. Yanling Karen Wang in the Roberts group for her guidance in my first year. I would like to express my thanks to present and past members of the Roberts group for discussions and help to my projects. I am happy to work and be friends with them. Last, a special thank goes to my family, my parents Xiaofei Wei and Lin Yuan, and my fiancé, Kai Hong, for their love, care and support in my life. iii Abbreviation A. fulgidus Archaeaoglubus fulgidus AHS adjacent hydrophobic site C2 protein kinase C conserved region 2 CD circular dichroism CMC critical micelle concentration CSA chemical shift anisotropy DAG diacylglycerol Dansyl-PE 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(5- dimethylamino-1-naphthalene-sulfonyl) DGP diglycerol phosphate D-I-1-P D-myo-inositol 1-phosphate diC6PI 1,2-dihexanoyl-sn-glycero-3-phospho-(1’-myo-inositol) 1,2-dihexanoyl-sn-glycero-3-phospho-(1’-myo-inositol-3’,4’- diC6PI(3,4)P2 bisphosphate) 1,2-dihexanoyl-sn-glycero-3-phospho-(1’-myo-inositol-4’,5’- diC6PI(4,5)P2 bisphosphate) diC7PC 1,2-diheptanoyl-sn-glycero-3-phosphocholine diC8PI 1,2-dioctanoyl-sn-glycero-3-phospho-(1’-myo-inositol) diC8PI(3)P 1,2-dioctanoyl-sn-glycero-3-phospho-(1’-myo-inositol-3’- phosphate) 1,2-dioctanoyl-sn-glycero-3-phospho-(1’-myo-inositol-3’,4’- diC8PI(3,4)P2 bisphosphate) 1,2-dioctanoyl-sn-glycero-3-phospho-(1’-myo-inositol-4’,5’- diC8PI(4,5)P2 bisphosphate) iv DIP di-myo-inositol-1,1’ phosphate 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol-3’,4’- DOPI(3,4)P2 bisphosphate) 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol-4’,5’- DOPI(4,5)P2 bisphosphate) DTT dithioreitol FBP fructose 1,6-bisphosphate G6P glucose-6-phosphate I(1,3,4,5)P4 or IP4 D-myo-inositol-1,3,4,5-tetraphosphate I(1,4,5)P3 or IP3 D-myo-inositol-1,4,5-triphosphate IMPase inositol monophosphatase IP inositol phosphate IPTG isopropyl-β-D-thiogalactopyranoside L-3,5-dd-diC8PI L-3,5-dideoxy-dioctanoyl-phosphatidylinositol L-I-1-P L-myo-inositol 1-phosphate LUV large unilamellar vesicle MTSL S-(2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl)methyl methanesulfonothioate NBP N-terminal PI(4,5)P2 binding patch NMR nuclear magnetic resonance PA phosphatidic acid PAGE polyacrylamide gel electrophoresis v PCR polymerase chain reaction PDZ post synaptic density protein, drosophila disc large tumor suppressor, and zonula occludens-1 protein PH pleckstrin homology PI phosphatidylinositol PI(3)P phosphatidylinositol 3-phosphate PI(3,4)P2 phosphatidylinositol 3,4-bisphosphate PI(3,4,5)P3 phosphatidylinositol 3,4,5-triphosphate PI(3,5)P2 phosphatidylinositol 3,5-bisphosphate PI(4,5)P2 phosphatidylinositol 4,5-bisphosphate PI3K phosphatidylinositol 3-phosphate kinase PLC phospholipase C POPC 1-palmitoyl-2-oleoyl-sn-glycero-phosphocholine PS phosphatidylserine PTEN phosphatase and tensin homolog deleted on chromosome ten Pyrene-PE 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(1- pyrenesulfonyl) SDS sodium dodecyl sulfate SUV small unilamellar vesicle TX-100 triton X-100 WT wild type vi Table of Contents Chapter 1 Introduction ................................................................................................... 1 1.1 Inositol and inositol metabolism .............................................................................. 2 1.2 Function of inositol phosphates in eukaryotes ......................................................... 6 1.3 Function of phosphoinositides in eukaryotes ........................................................... 7 1.3.1 Phosphatidylinositol monophosphates: PI(3)P, PI(4)P and PI(5)P ...................... 8 1.3.2 Phosphatidylinositol bisphosphates: PI(3,5)P2, PI(4,5)P2 and PI(3,4)P2 ........... 10 1.3.3 PI(3,4,5)P3 and the PI 3-kinase (PI3K) signaling pathway ................................ 12 1.4 PTEN...................................................................................................................... 15 1.4.1 Substrate specificity ........................................................................................... 16 1.4.2 PTEN structure ................................................................................................... 17 1.4.3 PTEN membrane binding and the role of the N-terminal loop .......................... 21 1.4.4 PTEN C-tail, regulation by phosphorylation...................................................... 27 1.4.5 Regulation of PTEN by other post-translational modification........................... 30 1.4.6 PDZ binding domain and regulation of PTEN by protein-protein
Load more

Recommended publications
  • Role and Significance of Sucrose-6-Phosphate Phosphatase in Regulating Sucrose Biosynthesis and Carbon Partitioning in Photosynthetic and Non-Photosynthetic Tissues
    Role and significance of sucrose-6-phosphate phosphatase in regulating sucrose biosynthesis and carbon partitioning in photosynthetic and non-photosynthetic tissues Dissertation zur Erlangung des akademischen Grades Doktor der Naturwissenschaften -Dr. rer. nat.- vorgelegt der Mathematisch-Naturwissenschaftlich-Technischen Fakultät (mathematisch-naturwissenschaftlicher Bereich) der Martin-Luther-Universität Halle-Wittenberg von Herrn Shuai Chen geb. am 29. 08. 1975 in Shandong, Volksrepublik China 1. Gutachter: Prof. Dr. Uwe Sonnewald 2. Gutachter: Prof. Dr. Klaus Humbeck Halle (Saale), den 29. August 2005 urn:nbn:de:gbv:3-000008943 [http://nbn-resolving.de/urn/resolver.pl?urn=nbn%3Ade%3Agbv%3A3-000008943] Contents Contents 1 Introduction 1 1.1 Sink and source concept 1 1.2 Carbon partitioning between starch- and sucrose-synthesis in source leaves 2 1.3 Sucrose synthesis in source leaves 4 1.4 Phloem loading and long-distance transport of sucrose 6 1.5 Sucrose unloading and metabolism in sink organs 7 1.6 Sink regulation of photosynthesis and sugar signalling 10 1.7 Reversed genetics approaches for the identification of metabolic control steps 13 1.8 Chemical-inducible expression of transgenes to study plant metabolism 15 1.9 Scientific aims of this work 17 2 Materials and methods 18 2.1 Chemicals, enzymes and other consumables 18 2.2 Plant materials and growth conditions 18 2.2.1 Nicotiana tabacum 18 2.2.2 Solanum tuberosum 18 2.3 DNA cloning procedures 19 2.4 Oligonucleotides and DNA Sequencing 19 2.5 E. coli strains and plasmids 19
    [Show full text]
  • The Action of the Phosphatases of Human Brain on Lipid Phosphate Esters by K
    J Neurol Neurosurg Psychiatry: first published as 10.1136/jnnp.19.1.12 on 1 February 1956. Downloaded from J. Neurol. Neurosurg. Psychiat., 1956, 19, 12 THE ACTION OF THE PHOSPHATASES OF HUMAN BRAIN ON LIPID PHOSPHATE ESTERS BY K. P. STRICKLAND*, R. H. S. THOMPSON, and G. R. WEBSTER From the Department of Chemical Pathology, Guy's Hospital Medical School, London, Much work, using both histochemical and therefore to study the action of the phosphatases in standard biochemical techniques, has been carried human brain on the " lipid phosphate esters out on the phosphatases of peripheral nerve. It is i.e., on the various monophosphate esters that occur known that this tissue contains both alkaline in the sphingomyelins, cephalins, and lecithins. In (Landow, Kabat, and Newman, 1942) and acid addition to ox- and 3-glycerophosphate we have phosphatases (Wolf, Kabat, and Newman, 1943), therefore used phosphoryl choline, phosphoryl and the changes in the levels of these enzymes in ethanolamine, phosphoryl serine, and inositol nerves undergoing Wallerian degeneration following monophosphate as substrates for the phospho- transection have been studied by several groups of monoesterases, and have measured their rates guest. Protected by copyright. of investigators (see Hollinger, Rossiter, and Upmalis, hydrolysis by brain preparations over the pH range 1952). 4*5 to 100. Phosphatase activity in brain was first demon- Plimmer and Burch (1937) had earlier reported strated by Kay (1928), and in 1934 Edlbacher, that phosphoryl choline and phosphoryl ethanol- Goldschmidt, and Schiiippi, using ox brain, showed amine are hydrolysed by the phosphatases of bone, that both acid and alkaline phosphatases are kidney, and intestine, but thepH at which the hydro- present in this tissue.
    [Show full text]
  • Exosome-Mediated MIR211 Modulates Tumor Microenvironment Via the DUSP6-ERK5 Axis and Contributes to BRAFV600E Inhibitor Resistan
    bioRxiv preprint doi: https://doi.org/10.1101/548818; this version posted February 13, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Exosome-mediated MIR211 modulates tumor microenvironment via the DUSP6-ERK5 axis and contributes to BRAFV600E inhibitor resistance in melanoma. Bongyong Lee1,3, Anupama Sahoo3, Junko Sawada1,3, John Marchica1,3, Sanjay Sahoo3, Fabiana I. A. L. Layng4, Darren Finlay4, Joseph Mazar5, Piyush Joshi1, Masanobu Komatsu1,3, Kristiina Vuori4, Garth Powis4, Petrus R. de Jong4, Animesh Ray6,7, and Ranjan J. Perera 1,2,3* 1 Department of Cancer and Blood Disorders Institute, Johns Hopkins All Children’s Hospital, St. Petersburg, FL 33701 USA 2 Department of Oncology, Sydney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287 USA 3 Sanford Burnham Prebys Medical Discovery Institute, Orlando, FL 32827 USA 4 Sanford Burnham Prebys Medical Discovery Institute, NCI-Designated Cancer Center, La Jolla, CA 92037, USA 5 Nemours Children Hospital, Orlando, FL 32827 USA 6 Keck Graduate Institute, Claremont CA 91711 USA, 7 Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125 USA. *Correspondence: Ranjan Perera Tel: 1-727-767-3491 Email: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/548818; this version posted February 13, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. ABSTRACT The microRNA MIR211 is an important regulator of melanoma tumor cell behavior.
    [Show full text]
  • Dual-Specificity Phosphatase 3 Deletion Protects Female, but Not
    Published August 28, 2017, doi:10.4049/jimmunol.1602092 The Journal of Immunology Dual-Specificity Phosphatase 3 Deletion Protects Female, but Not Male, Mice from Endotoxemia-Induced and Polymicrobial-Induced Septic Shock Maud M. Vandereyken,*,1 Pratibha Singh,*,1 Caroline P. Wathieu,* Sophie Jacques,* Tinatin Zurashvilli,* Lien Dejager,†,‡ Mathieu Amand,* Lucia Musumeci,* Maneesh Singh,* Michel P. Moutschen,* Claude R. F. Libert,†,‡ and Souad Rahmouni* Dual-specificity phosphatase 3 (DUSP3) is a small phosphatase with poorly known physiological functions and for which only a few substrates are known. Using knockout mice, we recently reported that DUSP3 deficiency confers resistance to endotoxin- and polymicrobial-induced septic shock. We showed that this protection was macrophage dependent. In this study, we further investigated the role of DUSP3 in sepsis tolerance and showed that the resistance is sex dependent. Using adoptive-transfer experiments and ovariectomized mice, we highlighted the role of female sex hormones in the phenotype. Indeed, in ovariec- tomized females and in male mice, the dominance of M2-like macrophages observed in DUSP32/2 female mice was reduced, suggesting a role for this cell subset in sepsis tolerance. At the molecular level, DUSP3 deletion was associated with estrogen- dependent decreased phosphorylation of ERK1/2 and Akt in peritoneal macrophages stimulated ex vivo by LPS. Our results demonstrate that estrogens may modulate M2-like responses during endotoxemia in a DUSP3-dependent manner. The Journal of Immunology, 2017, 199: 000–000. epsis and septic shock are complex clinical syndromes that ally, death (4). Sepsis occurrence and outcome depend on arise when the local body response to pathogens becomes pathogen characteristics, as well as on risk factors, such as age S systemic and injures its own tissues and organs (1).
    [Show full text]
  • Exploring Metagenomic Enzymes: a Novel Esterase Useful for Short-Chain Ester Synthesis
    catalysts Article Exploring Metagenomic Enzymes: A Novel Esterase Useful for Short-Chain Ester Synthesis 1,2, 1,2, 1 Thaís Carvalho Maester y , Mariana Rangel Pereira z, Aliandra M. Gibertoni Malaman , Janaina Pires Borges 3,Pâmela Aparecida Maldaner Pereira 1 and Eliana G. M. Lemos 1,* 1 Department of Technology, São Paulo State University (UNESP), Jaboticabal, SP 14884-900, Brazil; [email protected] (T.C.M.); [email protected] (M.R.P.); [email protected] (A.M.G.M.); [email protected] (P.A.M.P.) 2 Institute of Biomedical Sciences (ICB III), University of São Paulo (USP), São Paulo, SP 05508-900, Brazil 3 Institute of Biosciences, Languages and Exact Sciences, Department of Chemistry and Environmental Sciences, São Paulo State University (UNESP), São José do Rio Preto, SP 15054-000, Brazil; [email protected] * Correspondence: [email protected]; Tel.: +55-16-3209-7409 Current address: Supera Innovation and Technology Park, Ecobiotech Company, Ribeirão Preto, y SP 14056-680, Brazil. Current address: Department of Biochemistry, University of Cambridge, Cambridge CB2 1TN, UK. z Received: 13 August 2020; Accepted: 24 August 2020; Published: 23 September 2020 Abstract: Enzyme-mediated esterification reactions can be a promising alternative to produce esters of commercial interest, replacing conventional chemical processes. The aim of this work was to verify the potential of an esterase for ester synthesis. For that, recombinant lipolytic enzyme EST5 was purified and presented higher activity at pH 7.5, 45 ◦C, with a Tm of 47 ◦C. Also, the enzyme remained at least 50% active at low temperatures and exhibited broad substrate specificity toward p-nitrophenol esters 1 1 with highest activity for p-nitrophenyl valerate with a Kcat/Km of 1533 s− mM− .
    [Show full text]
  • The Regulatory Roles of Phosphatases in Cancer
    Oncogene (2014) 33, 939–953 & 2014 Macmillan Publishers Limited All rights reserved 0950-9232/14 www.nature.com/onc REVIEW The regulatory roles of phosphatases in cancer J Stebbing1, LC Lit1, H Zhang, RS Darrington, O Melaiu, B Rudraraju and G Giamas The relevance of potentially reversible post-translational modifications required for controlling cellular processes in cancer is one of the most thriving arenas of cellular and molecular biology. Any alteration in the balanced equilibrium between kinases and phosphatases may result in development and progression of various diseases, including different types of cancer, though phosphatases are relatively under-studied. Loss of phosphatases such as PTEN (phosphatase and tensin homologue deleted on chromosome 10), a known tumour suppressor, across tumour types lends credence to the development of phosphatidylinositol 3--kinase inhibitors alongside the use of phosphatase expression as a biomarker, though phase 3 trial data are lacking. In this review, we give an updated report on phosphatase dysregulation linked to organ-specific malignancies. Oncogene (2014) 33, 939–953; doi:10.1038/onc.2013.80; published online 18 March 2013 Keywords: cancer; phosphatases; solid tumours GASTROINTESTINAL MALIGNANCIES abs in sera were significantly associated with poor survival in Oesophageal cancer advanced ESCC, suggesting that they may have a clinical utility in Loss of PTEN (phosphatase and tensin homologue deleted on ESCC screening and diagnosis.5 chromosome 10) expression in oesophageal cancer is frequent, Cao et al.6 investigated the role of protein tyrosine phosphatase, among other gene alterations characterizing this disease. Zhou non-receptor type 12 (PTPN12) in ESCC and showed that PTPN12 et al.1 found that overexpression of PTEN suppresses growth and protein expression is higher in normal para-cancerous tissues than induces apoptosis in oesophageal cancer cell lines, through in 20 ESCC tissues.
    [Show full text]
  • Association of Variation in the Sugarcane Transcriptome with Sugar Content Prathima P
    Thirugnanasambandam et al. BMC Genomics (2017) 18:909 DOI 10.1186/s12864-017-4302-5 RESEARCH ARTICLE Open Access Association of variation in the sugarcane transcriptome with sugar content Prathima P. Thirugnanasambandam1,2†, Nam V. Hoang1,3†, Agnelo Furtado1, Frederick C. Botha4 and Robert J. Henry1,5* Abstract Background: Sugarcane is a major crop of the tropics cultivated mainly for its high sucrose content. The crop is genetically less explored due to its complex polyploid genome. Sucrose synthesis and accumulation are complex processes influenced by physiological, biochemical and genetic factors, and the growth environment. The recent focus on the crop for fibre and biofuel has led to a renewed interest on understanding the molecular basis of sucrose and biomass traits. This transcriptome study aimed to identify genes that are associated with and differentially regulated during sucrose synthesis and accumulation in the mature stage of sugarcane. Patterns of gene expression in high and low sugar genotypes as well as mature and immature culm tissues were studied using RNA-Seq of culm transcriptomes. Results: In this study, 28 RNA-Seq libraries from 14 genotypes of sugarcane differing in their sucrose content were used for studying the transcriptional basis of sucrose accumulation. Differential gene expression studies were performed using SoGI (Saccharum officinarum Gene Index, 3.0), SAS (sugarcane assembled sequences) of sugarcane EST database (SUCEST) and SUGIT, a sugarcane Iso-Seq transcriptome database. In total, about 34,476 genes were found to be differentially expressed between high and low sugar genotypes with the SoGI database, 20,487 genes with the SAS database and 18,543 genes with the SUGIT database at FDR < 0.01, using the Baggerley’s test.
    [Show full text]
  • Macrophage DUSP3 Genetic Deletion Confers M2-Like
    DUSP3 Genetic Deletion Confers M2-like Macrophage−Dependent Tolerance to Septic Shock This information is current as Pratibha Singh, Lien Dejager, Mathieu Amand, Emilie of September 27, 2021. Theatre, Maud Vandereyken, Tinatin Zurashvili, Maneesh Singh, Matthias Mack, Steven Timmermans, Lucia Musumeci, Emmanuel Dejardin, Tomas Mustelin, Jo A. Van Ginderachter, Michel Moutschen, Cécile Oury, Claude Libert and Souad Rahmouni Downloaded from J Immunol 2015; 194:4951-4962; Prepublished online 15 April 2015; doi: 10.4049/jimmunol.1402431 http://www.jimmunol.org/content/194/10/4951 http://www.jimmunol.org/ References This article cites 35 articles, 9 of which you can access for free at: http://www.jimmunol.org/content/194/10/4951.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 27, 2021 • No Triage! Every submission reviewed by practicing scientists • 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 © 2015 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology DUSP3 Genetic Deletion Confers M2-like Macrophage–Dependent Tolerance to Septic Shock Pratibha Singh,*,1 Lien Dejager,†,‡,1 Mathieu Amand,*,1 Emilie Theatre,x Maud Vandereyken,* Tinatin Zurashvili,* Maneesh Singh,* Matthias Mack,{ Steven Timmermans,†,‡ Lucia Musumeci,* Emmanuel Dejardin,‖ Tomas Mustelin,#,** Jo A.
    [Show full text]
  • Changes in Phosphatidylinositol Metabolism in Response to Hyperosmotic Stress in Daucus Carota 1. Cells Grown in Suspension Culture'
    Plant Physiol. (1993) 103: 637-647 Changes in Phosphatidylinositol Metabolism in Response to Hyperosmotic Stress in Daucus carota 1. Cells Grown in Suspension Culture' Myeon H. Cho, Stephen B. Shears, and Wendy F. BOSS* Department of Botany, North Carolina State University, Raleigh, North Carolina 27695-761 2 (M.H.C., W.F.B.); and Laboratory of Cellular and Molecular Pharmacology, National lnstitute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709 (S.B.S) in the Samanea saman pulvini is there evidence for stimulus- Carrot (Daucus carota L.) cells plasmolyzed within 30 s after mediated turnover of polyphosphorylated inositol phospho- adding sorbitol to increase the osmotic strength of the medium lipids and inositol phosphates within the rapid time scale from 0.2 to 0.4 or 0.6 osmolal. However, there was no significant seen in animal cells (Morse et al., 1987). Although results change in the polyphosphorylated inositol phospholipids or inositol from in vitro studies and from IP3 microinjection in vivo have phosphates or in inositol phospholipid metabolism within 30 s of shown that Ir3 can release Ca2+from internal stores such as imposing the hyperosmotic stress. Maximum changes in phospha- vacuoles and tonoplast vesicles (Drcibak and Ferguson, 1985; tidylinositol 4-monophosphate (PIP) metabolism were detected at 5 min, at which time the cells appeared to adjust to the change in Schumaker and Sze, 1987; Ranjeva et al., 1988; Gilroy et al., osmoticum. There was a 30% decrease in [3H]inositol-labeledPIP. 1990), the complete pathway from a stimulus to Ca2+release The specific activity of enzymes involved in the metabolism of the has yet to be demonstrated in higher plants.
    [Show full text]
  • Human Induced Pluripotent Stem Cell–Derived Podocytes Mature Into Vascularized Glomeruli Upon Experimental Transplantation
    BASIC RESEARCH www.jasn.org Human Induced Pluripotent Stem Cell–Derived Podocytes Mature into Vascularized Glomeruli upon Experimental Transplantation † Sazia Sharmin,* Atsuhiro Taguchi,* Yusuke Kaku,* Yasuhiro Yoshimura,* Tomoko Ohmori,* ‡ † ‡ Tetsushi Sakuma, Masashi Mukoyama, Takashi Yamamoto, Hidetake Kurihara,§ and | Ryuichi Nishinakamura* *Department of Kidney Development, Institute of Molecular Embryology and Genetics, and †Department of Nephrology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan; ‡Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Hiroshima, Japan; §Division of Anatomy, Juntendo University School of Medicine, Tokyo, Japan; and |Japan Science and Technology Agency, CREST, Kumamoto, Japan ABSTRACT Glomerular podocytes express proteins, such as nephrin, that constitute the slit diaphragm, thereby contributing to the filtration process in the kidney. Glomerular development has been analyzed mainly in mice, whereas analysis of human kidney development has been minimal because of limited access to embryonic kidneys. We previously reported the induction of three-dimensional primordial glomeruli from human induced pluripotent stem (iPS) cells. Here, using transcription activator–like effector nuclease-mediated homologous recombination, we generated human iPS cell lines that express green fluorescent protein (GFP) in the NPHS1 locus, which encodes nephrin, and we show that GFP expression facilitated accurate visualization of nephrin-positive podocyte formation in
    [Show full text]
  • Some Ultrastructural and Enzymatic Effects of Water Stress in Cotton (Gossypium Hirsutum L.) Leaves (Acid Phosphatase/Acid Lipase/Alkaline Lipase)
    Proc. Nat. Acad. Sci. USA Vol. 71, No. 8, pp. 3243-3247, August 1974 Some Ultrastructural and Enzymatic Effects of Water Stress in Cotton (Gossypium hirsutum L.) Leaves (acid phosphatase/acid lipase/alkaline lipase) JORGE VIEIRA DA SILVA*, AUBREY W. NAYLOR, AND PAUL J. KRAMER Department of Botany, Duke University, Durham, North Carolina 27706 Contributed by Paul J. Kramer, May 30, 1974 ABSTRACT Water stress induced by floating discs cut boxylation of glycine occurs after lipase treatment of mito- from cotton leaves (Gossypium hirsutum L. cultivar chondria Stoneville) on a polyethylene glycol solution (water poten- (23). tial, -10 bars) was associated with marked alteration of Results, thus far obtained by indirect means, support the ultrastructural organization of both chloroplasts and hypothesis that water stress in drought sensitive species leads mitochondria. Ultrastructural organization of chloro- to hydrolytic activity that degrades not only storage products plasts was sometimes almost completely destroyed; per- but the structural framework of organelles such as ribosomes, oxisomes seemed not to be affected; and chloroplast ribosomes disappeared. Also accompanying water stress chloroplasts, and mitochondria. Ultrastructural and micro- was a sharp increase in activity of acid phosphatase chemical evidence is reported here that such deterioration [orthoplhosphoric-monoester phosplhohydrolase (acid opti- occurs in cotton (Gossypium hirsutum L. cv. Stoneville) during mum), EC 3.1.3.2], and acid and alkaline lipase [glycerol water stress. ester hydrolase EC 3.1.1.3] within chloroplasts. Only acid lipase activity was detected inside mitochondria of stressed MATERIALS AND METHODS discs. These alterations in cell organization and enzy- mology may account for at least part of the previously Cotton plants (Gossypium hirsutum L.
    [Show full text]
  • Human Erythrocyte Acetylcholinesterase
    Pediat. Res. 7: 204-214 (1973) A Review: Human Erythrocyte Acetylcholinesterase FRITZ HERZ[I241 AND EUGENE KAPLAN Departments of Pediatrics, Sinai Hospital, and the Johns Hopkins University School of Medicine, Baltimore, Maryland, USA Introduction that this enzyme was an esterase, hence the term "choline esterase" was coined [100]. Further studies In recent years the erythrocyte membrane has received established that more than one type of cholinesterase considerable attention by many investigators. Numer- occurs in the animal body, differing in substrate ous reviews on the composition [21, 111], immunologic specificity and in other properties. Alles and Hawes [85, 116] and rheologic [65] properties, permeability [1] compared the cholinesterase of human erythro- [73], active transport [99], and molecular organiza- cytes with that of human serum and found that, tion [109, 113, 117], attest to this interest. Although although both enzymes hydrolyzed acetyl-a-methyl- many studies relating to membrane enzymes have ap- choline, only the erythrocyte cholinesterase could peared, systematic reviews of this area are limited. hydrolyze acetyl-yg-methylcholine and the two dia- More than a dozen enzymes have been recognized in stereomeric acetyl-«: /3-dimethylcholines. These dif- the membrane of the human erythrocyte, although ferences have been used to delineate the two main changes in activity associated with pathologic condi- types of cholinesterase: (1) acetylcholinesterase, or true, tions are found regularly only with acetylcholinesterase specific, E-type cholinesterase (acetylcholine acetyl- (EC. 3.1.1.7). Although the physiologic functions of hydrolase, EC. 3.1.1.7) and (2) cholinesterase or erythrocyte acetylcholinesterase remain obscure, the pseudo, nonspecific, s-type cholinesterase (acylcholine location of this enzyme at or near the cell surface gives acylhydrolase, EC.
    [Show full text]