DOCSLIB.ORG

Macrophage-Activating Factors Glycoprotein Are Distinct

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Macrophage-Activating Factors Glycoprotein Are Distinct The Glycosyl-Inositol-Phosphate and Dimyristoylglycerol Moieties of the Glycosylphosphatidylinositol Anchor of the Trypanosome Variant-Specific Surface This information is current as Glycoprotein Are Distinct of September 27, 2021. Macrophage-Activating Factors Stefan Magez, Benoi?t Stijlemans, Magdalena Radwanska, Etienne Pays, Michael A. J. Ferguson and Patrick De Baetselier J Immunol 1998; 160:1949-1956; ; Downloaded from http://www.jimmunol.org/content/160/4/1949 References This article cites 40 articles, 21 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/160/4/1949.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 27, 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 © 1998 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Glycosyl-Inositol-Phosphate and Dimyristoylglycerol Moieties of the Glycosylphosphatidylinositol Anchor of the Trypanosome Variant-Specific Surface Glycoprotein Are Distinct Macrophage-Activating Factors1 Stefan Magez,2* Benoıˆt Stijlemans,* Magdalena Radwanska,† Etienne Pays,† Michael A. J. Ferguson,‡ and Patrick De Baetselier* The TNF-a-inducing capacity of different trypanosome components was analyzed in vitro, using as indicator cells a macrophage cell line (2C11/12) or peritoneal exudate cells from LPS-resistant C3H/HeJ mice and LPS-sensitive C3H/HeN mice. The variant- specific surface glycoprotein (VSG) was identified as the major TNF-a-inducing component present in trypanosome-soluble extracts. Both soluble (sVSG) and membrane-bound VSG (mfVSG) were shown to manifest similar TNF-a-inducing capacities, Downloaded from indicating that the dimyristoylglycerol (DMG) compound of the mfVSG anchor was not required for TNF-a triggering. Detailed analysis indicated that the glycosyl-inositol-phosphate (GIP) moiety was responsible for the TNF-a-inducing activity of VSG and that the presence of the GIP-associated galactose side chain was essential for optimal TNF-a production. Furthermore, the results showed that the responsiveness of macrophages toward the TNF-a-inducing activity of VSG was strictly dependent on the activation state of the macrophages, since resident macrophages required IFN-g preactivation to become responsive. Comparative analysis of the ability of both forms of VSG to activate macrophages revealed that mfVSG but not sVSG stimulates macrophages http://www.jimmunol.org/ toward IL-1a secretion and acquisition of LPS responsiveness. The priming activity of mfVSG toward LPS responsiveness was also demonstrated in vivo and may be relevant during trypanosome infections, since Trypanosoma brucei-infected mice became gradually LPS-hypersensitive during the course of infection. Collectively, the VSG of trypanosomes encompasses two distinct macrophage-activating components: while the GIP moiety of sVSG mediates TNF-a induction, the DMG compound of the mfVSG anchor contributes to IL-1a induction and LPS sensitization. The Journal of Immunology, 1998, 160: 1949–1956. umor necrosis factor a is a pleiotropic cytokine, produced In several of these infections, TNF-a plays a bidirectional role, mainly by activated macrophages (1). Although its name influencing both parasite and host (8). In the case of trypanosome by guest on September 27, 2021 T is derived from its capacity to cause necrosis of certain infections, TNF-a seems to be involved in the neuropathology of parenchymal organs and tumors, the molecule was initially also sleeping sickness, and a correlation has been described between isolated as a factor named “cachectin,” responsible for systemic the disease severity in human African trypanosomiasis and high suppression of lipoprotein lipase activity in trypanosome-induced serum levels of the cytokine (9). Furthermore, a possible associ- cachexia (2, 3). As TNF-a is a main mediator in inflammatory ation between TNF-a production by monocytes from trypano- responses, its influence in a number of infectious disease has been some-infected cattle and the severity of disease-associated anemia studied. Among other effects, TNF-a was found to be involved in has been documented (10). Using mouse models, the immunopa- the pathology of several parasitic diseases, including trypanoso- thology of experimental African sleeping sickness has been linked miasis (4), Chagas’ disease (5), leishmaniasis (6), and malaria (7). to TNF-a-mRNA detection in the brain of Trypanosoma brucei- infected mice (11), and a role for TNF-a involvement in trypano- somiasis-associated immunosuppression has been put forward a *Laboratory of Cellular Immunology, Flanders Interuniversity Institute for Biotech- (12). Finally, a direct involvement of TNF- in trypanosomiasis nology, Free University of Brussels (Vrije Universiteit Brussel), and †Department of control and parasite growth was documented in vivo (4, 13), and a Molecular Biology, Free University of Brussels (Universite´Libre de Bruxelles), Brus- trypanolytic effect of TNF-a on T. brucei was confirmed in vitro sels, Belgium; and ‡Department of Biochemistry, The University Dundee, Scotland, United Kingdom (14–16). Information concerning the trypanosome-derived fac- a Received for publication April 17, 1997. Accepted for publication October 31, 1997. tor(s) that may be responsible for the induction of TNF- The costs of publication of this article were defrayed in part by the payment of page production by host cells is scarce. In analogy with plasmodia-as- charges. This article must therefore be hereby marked advertisement in accordance sociated TNF-a induction, trypanosomiasis-associated TNF-a in- with 18 U.S.C. Section 1734 solely to indicate this fact. duction has been suggested to be mediated by the glycosylphos- 1 This project has been funded by the UNDP/World Bank/WHO Special Programme phatidylinositol (GPI)3 anchor on the variant-specific surface for Research and Training in Tropical Diseases, The Belgian National Fund for Sci- entific Research (FWO-No. 6.0325.95 and FNRS-No. 3.4502.96), the Flemish Gov- glycoprotein (VSG) of the trypanosome (17). VSG is the crucial ernment (Vlaams Actieprogramma Biotechnologie-VLAB), the European Commu- molecule involved in the escape of the host immune response, as nity (Cost contract No. TS3-CT94-0293), and the Communaute´Franc¸aise de Belgique 107 identical VSGs cover each trypanosome and form a dense coat (ARC 94/99-189). This project was performed within the framework of an Interuni- versity Attraction Pole Programme, financed by the Belgian state government (Dien- around each cell (18, 19). The mechanism of immune escape is sten van de Eerste Minister-Federale diensten voor wetenschappelijke, technische en culturele aangelegenheden). S.M. is a Postdoctoral Research Fellow of the Foundation of Scientific Research-Flanders (FWO). 3 Abbreviations used in this paper: GPI, glycosylphosphatidylinositol; DMG, dimyr- 2 Address correspondence and reprint requests to Stefan Magez, Eenheid CIMM istoylglycerol; GIP, glycosyl-inositol-phosphate; VSG, variant-specific surface gly- (IMOL 2), Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit coprotein; sVSG, soluble VSG; mfVSG, membrane form VSG; PEC, peritoneal ex- Brussel, Paardenstraat 65, 1640 Sint Genesius Rode, Belgium. udate cell; TLCK, Na-( p-tosyl)lysine chloromethyl ketone. Copyright © 1998 by The American Association of Immunologists 0022-1767/98/$02.00 1950 VSG-MEDIATED INDUCTION OF CYTOKINE PRODUCTION based on a regular switch in the expression of VSG variants, jeop- recombinant N-glycosidase F from Flavobacterium meningosepticum, ardizing the induction of an effective Ab response against the par- a-galactosidase from green coffee beans, Pronase from Streptomyces gri- asite. Interestingly, in response to environmental stress, trypano- seus, Proteinase K from Tritirachium album, and the protease inhibitors Pefabloc SC, PMSF, and TLCK. All enzymes and inhibitors were used somes are capable of liberating their VSG through a VSG lipase according to the manufacturer’s instructions. (20). This enzyme cleaves the GPI anchor, leaving the dimyris- N-Glycosidase F digestion was performed in PBS using 5 U/ml enzyme toylglycerol (DMG) compound of the GPI anchored in the mem- (24 h/37°C). brane, and releasing the glycosyl-inositol-phosphate (GIP)-VSG a-Galactosidase digestion of VSG was performed in 0.1 M sodium acetate buffer, pH 5.0, using 50 U/ml enzyme (2 3 18 h/37°C). part (21). In this report, we compare the macrophage-activating Proteinase K digestion was performed in PBS, pH 8.0, using 5 U/ml capacity, analyzed in terms of cytokine secretion (TNF-a/IL-1a, enzyme (24 h/37°C). IL-6, IL-10, and IL-12) and induction of LPS hyper-responsive- Pefabloc SC was used when indicated (1 mM final concentration) to ness, of the released soluble VSG (sVSG) and the membrane- prevent proteolysis during lysate preparation
Load more

Recommended publications
  • Snapshot: Inositol Phosphates Ace J
    SnapShot: Inositol Phosphates Ace J. Hatch and John D. York HHMI, Pharmacology and Cancer Biology, Biochemistry, Duke University, Durham, NC 27710, USA PLC-dependent IP code GPCR RTK O O O O O O 5-PP-IP4 IP4 5-IP7 O O O O O O PIP2 O IP6K O IP6K O VIP1 O O O 2 O ITPK1 O 13 O PLC 2 O O O O O O O O 4 6 13 O 5 IP3 IPMK IP4 IPMK IP5 IPK1 IP6 1,5-IP8 4 6 O O 5 O O O O O O O O ENZYMES O O O O O O YEAST MAMMALIAN IP3K VIP1 IP6K IPMK PLC1 PLCβ, γ, δ, ε, ζ, η O - IP3KA, B, C - ITPK1 (IP56K) O O O O O O O O IPK2(ARG82) IPMK (IPK2) IP4 IP3 IP4 1-IP7 IPK1 IPK1 (IP5K) INPP5 ITPK1 KCS1 IP6K1, 2, 3 O O O O O O VIP1 VIP1, 2 (PPIP5K1, 2) O O Ion channels Phosphate sensing Transcription Cl- Abundant phosphate MCM1 ARG80 CIC3 P PLASMA MEMBRANE - Pho80 Cl channel Pho4 Kinase Kinase Assembly Pho85 independent CYTOPLASM activity 2 O PIP2 Pho81 13 CYTOPLASM NUCLEUS IPK2 ARG81 4 6 Phosphate starvation MCM1-ArgR O 5 O complex O O IP4 O O O O O O O 1-IP7 Kinase Activation dependent IP3 O O Transcription O O O activated Pho80 IP4 O X Pho4 O O Pho85 Kinase activity IP receptor blocked O 3 ENDOPLASMIC Pho81 RETICULUM Ca2+ CYTOPLASM NUCLEUS NUCLEUS mRNA export and translation Insulin secretion and AKT Embryonic development Translation termination Effects of IP kinase deficiency O IPMK (IPK2): Multiple defects, death by embryonic day 10 (mice) O O Insulin IPK1: Cillia are shortened and immotile IP6 AKT resistance causing patterning defects (zebrash) O O Multiple defects, death by Ribosome O embryonic day 8.5 (mice) GleI eRF1 Insulin GSK3β Dbp5 ITPK1 (IP56K): Neural tube
    [Show full text]
  • ITPK1 Mediates the Lipid-Independent Synthesis of Inositol Phosphates Controlled by Metabolism
    ITPK1 mediates the lipid-independent synthesis of inositol phosphates controlled by metabolism Yann Desfougèresa, Miranda S. C. Wilsona, Debabrata Lahaa, Gregory J. Millerb, and Adolfo Saiardia,1 aMedical Research Council Laboratory for Molecular Cell Biology, University College London, WC1E 6BT London, United Kingdom; and bDepartment of Chemistry, The Catholic University of America, Washington, DC 20064 Edited by Solomon H. Snyder, The Johns Hopkins University School of Medicine, Baltimore, MD, and approved October 25, 2019 (received for review July 3, 2019) Inositol phosphates (IPs) comprise a network of phosphorylated The potential pathways to IP6-7-8 synthesis have a profound molecules that play multiple signaling roles in eukaryotes. IPs impact on how we interpret their signaling roles. If they are synthesis is believed to originate with IP3 generated from PIP2 by synthesized from IP3, their function has evolved in relation to phospholipase C (PLC). Here, we report that in mammalian cells lipid-dependent PLC and/or calcium signaling. This constraint is PLC-generated IPs are rapidly recycled to inositol, and uncover the relieved if IP6 synthesis occurs lipid-independently, originating enzymology behind an alternative “soluble” route to synthesis of directly from inositol, a “soluble” pathway. In yeast, which do not IPs. Inositol tetrakisphosphate 1-kinase 1 (ITPK1)—found in Asgard have IP3-regulated calcium signaling, IP6 synthesis strictly depends archaea, social amoeba, plants, and animals—phosphorylates on the PLC-generated IP3 (Figs. 1A and 2D). The higher com- I(3)P1 originating from glucose-6-phosphate, and I(1)P1 generated plexity of other eukaryotes might allow different pathways for IPs from sphingolipids, to enable synthesis of IP6.
    [Show full text]
  • Glycosylphosphatidylinositol Anchors from Galactomannan and GPI-Anchored Protein Are Synthesized by Distinct Pathways in Aspergillus Fumigatus
    Journal of Fungi Article Glycosylphosphatidylinositol Anchors from Galactomannan and GPI-Anchored Protein Are Synthesized by Distinct Pathways in Aspergillus fumigatus Jizhou Li 1, Isabelle Mouyna 1, Christine Henry 1, Frédérique Moyrand 2, Christian Malosse 3, Julia Chamot-Rooke 3, Guilhem Janbon 2, Jean-Paul Latgé 1 and Thierry Fontaine 1,* 1 Unité des Aspergillus, 25 rue du Docteur Roux, Institut Pasteur, 25 rue du Docteur Roux, 75015 Paris, France; [email protected] (J.L.); [email protected] (I.M.); [email protected] (C.H.); [email protected] (J.-P.L.) 2 Unité de Biologie des ARN des Pathogènes Fongiques, Institut Pasteur, 25 rue du Docteur Roux, 75015 Paris, France; [email protected] (F.M.); [email protected] (G.J.) 3 Unité de Spectrométrie de Masse pour la Biologie, Institut Pasteur, CNRS USR 2000, 28 rue du Docteur Roux, 75015 Paris, France; [email protected] (C.M.); [email protected] (J.C.-R.) * Correspondence: [email protected]; Tel.: +33-145-688-358 Received: 8 December 2017; Accepted: 19 January 2018; Published: 2 Febuary 2018 Abstract: Glycosylphosphatidylinositols (GPIs) are lipid anchors allowing the exposure of proteins at the outer layer of the plasma membrane. In fungi, a number of GPI-anchored proteins (GPI-APs) are involved in the remodeling of the cell wall polymers. GPIs follow a specific biosynthetic pathway in the endoplasmic reticulum. After the transfer of the protein onto the GPI-anchor, a lipid remodeling occurs to substitute the diacylglycerol moiety by a ceramide. In addition to GPI-APs, A. fumigatus produces a GPI-anchored polysaccharide, the galactomannan (GM), that remains unique in the fungal kingdom.
    [Show full text]
  • Dephosphorylation of 2,3-Bisphosphoglycerate by MIPP Expands the Regulatory Capacity of the Rapoport–Luebering Glycolytic Shunt
    Dephosphorylation of 2,3-bisphosphoglycerate by MIPP expands the regulatory capacity of the Rapoport–Luebering glycolytic shunt Jaiesoon Cho*†, Jason S. King‡§, Xun Qian*, Adrian J. Harwood‡, and Stephen B. Shears*¶ *Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Social Services, P.O. Box 12233, Research Triangle Park, NC 27709; and ‡Cardiff School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3US, United Kingdom Edited by Helen M. Ranney, University of California at San Diego, La Jolla, CA, and approved March 6, 2008 (received for review November 21, 2007) The Rapoport–Luebering glycolytic bypass comprises evolutionar- majority of mammals. By preferentially binding to deoxyhemo- ily conserved reactions that generate and dephosphorylate 2,3- globin, 2,3-BPG facilitates oxygen release from the erythrocyte bisphosphoglycerate (2,3-BPG). For >30 years, these reactions have to the surrounding tissues (6, 7). Thus, the regulation of eryth- been considered the responsibility of a single enzyme, the 2,3-BPG rocyte 2,3-BPG levels is key to efficiently meeting tissue oxygen synthase/2-phosphatase (BPGM). Here, we show that Dictyosteli- demands while also providing an important physiological adap- um, birds, and mammals contain an additional 2,3-BPG phospha- tation to oxygen deprivation (8), including that which occurs at tase that, unlike BPGM, removes the 3-phosphate. This discovery high altitude (9) or during postoperative anemia (10). reveals that the glycolytic pathway can bypass the formation of Despite the importance of carefully regulating 2,3-BPG turn- 3-phosphoglycerate, which is a precursor for serine biosynthesis over, little is known about how this might be achieved.
    [Show full text]
  • Structural Basis for Phosphatidylinositol-Phosphate Biosynthesis
    ARTICLE Received 8 Jun 2015 | Accepted 29 Aug 2015 | Published 16 Oct 2015 DOI: 10.1038/ncomms9505 OPEN Structural basis for phosphatidylinositol-phosphate biosynthesis Oliver B. Clarke1,*, David Tomasek2,*, Carla D. Jorge3,*, Meagan Belcher Dufrisne2, Minah Kim2, Surajit Banerjee4, Kanagalaghatta R. Rajashankar4, Lawrence Shapiro1, Wayne A. Hendrickson1, Helena Santos3 & Filippo Mancia2 Phosphatidylinositol is critical for intracellular signalling and anchoring of carbohydrates and proteins to outer cellular membranes. The defining step in phosphatidylinositol biosynthesis is catalysed by CDP-alcohol phosphotransferases, transmembrane enzymes that use CDP-diacylglycerol as donor substrate for this reaction, and either inositol in eukaryotes or inositol phosphate in prokaryotes as the acceptor alcohol. Here we report the structures of a related enzyme, the phosphatidylinositol-phosphate synthase from Renibacterium salmoninarum, with and without bound CDP-diacylglycerol to 3.6 and 2.5 Å resolution, respectively. These structures reveal the location of the acceptor site, and the molecular determinants of substrate specificity and catalysis. Functional characterization of the 40%-identical ortholog from Mycobacterium tuberculosis, a potential target for the development of novel anti-tuberculosis drugs, supports the proposed mechanism of substrate binding and catalysis. This work therefore provides a structural and functional framework to understand the mechanism of phosphatidylinositol-phosphate biosynthesis. 1 Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA. 2 Department of Physiology and Cellular Biophysics, Columbia University, New York, NY 10032, USA. 3 Biology Division, Instituto de Tecnologia Quı´mica e Biolo´gica, Universidade Nova de Lisboa, Avenida da Repu´blica-EAN, 2780-157 Oeiras, Portugal. 4 NE-CATand Department of Chemistry and Chemical Biology, Cornell University, Argonne National Laboratory, Argonne, IL 60439, USA.
    [Show full text]
  • Glycosyl-Phosphatidylinositol/Inositol Phosphoglycan
    Proc. Natl. Acad. Sci. USA Vol. 88, pp. 8016-8019, September 1991 Biochemistry Glycosyl-phosphatidylinositol/inositol phosphoglycan: A signaling system for the low-affinity nerve growth factor receptor (development/inner ear/cochleovestibular ganglion/ant-ostol phosphoglycan antibody) JUAN REPRESA*, MATfAS A. AVILAt, CRISTINA MINERf, FERNANDO GIRALDEZt, GUILLERMO ROMERO§, ROSA CLEMENTEt, JOSE M. MATOt, AND ISABEL VARELA-NIETOt¶ *Departamento Ciencias Morfol6gicas and *Departamento Bioqutmica, Biologfa Molecular y Fisiologfa, Facultad de Medicina, Universidad de Valladolid, 47005 Valladolid, Spain; §Department of Pharmacology, University of Virginia, Charlottesville, VA 22908; and tInstituto de Investigaciones Biomddicas, Consejo Superior de Investigaciones Cientfficas and Departamento Bioqufmica, Universidad Aut6noma de Madrid, Arturo Duperier 4, 28029 Madrid, Spain Communicated by Sidney Udenfriend, June 7, 1991 (received for review April 15, 1991) ABSTRACT Nerve growth factor (NGF) exerts a variety of that IPG would be conserved for some of the developmental actions during embryonic development. At the early stages of actions of insulin and NGF, which could use a common inner ear development, NGF stimulates cell proliferation, an signaling pathway, shared perhaps with other related growth effect mediated through low-affinity receptors. We have stud- factors, to regulate cell growth. The present work provides ied the possibility that the glycosyl-phosphatidylinositol/ further support for the involvement of this glycosyl-PtdIns/ inositol phosphoglycan (glycosyl-Ptdlns/IPG) system is in- IPG pathway in transducing the mitogenic effects of NGF on volved in transmitting this NGF signal. Endogenous glycosyl- the early developing inner ear by showing the following PtdIns was characterized in extracts of cochleovestibular gan- results: (i) the presence of endogenous glycosyl-PtdIns and glia (CVGs) that incorporated [3Hglucosamine, [Hjglactose, IPG, the latter with strong mitogenic activity; (it) the ability [3lH]myristic acid, and PH]palmitic acid.
    [Show full text]
  • Phosphatidylinositol 4,5-Bisphosphate Modifies Tubulin
    The Journal of Neuroscience, March 1, 2002, 22(5):1668–1678 Phosphatidylinositol 4,5-Bisphosphate Modifies Tubulin ␤ Participation in Phospholipase C 1 Signaling Juliana S. Popova,1 Arin K. Greene,1 Jia Wang,1 and Mark M. Rasenick1,2 Departments of 1Physiology and Biophysics and 2Psychiatry, University of Illinois at Chicago, College of Medicine, Chicago, Illinois 60612-7342 Tubulin forms the microtubule and regulates certain G-protein- the plasma membrane was demonstrated with confocal laser mediated signaling pathways. Both functions rely on the GTP- immunofluorescence microscopy. Although tubulin bound to ␣ ␣ ␤ binding properties of tubulin. Signal transduction through G q- both G q and PLC 1 , PIP2 facilitated the interaction between ␤ ␤ ␤ ␣ regulated phospholipase C 1 (PLC 1 ) is activated by tubulin tubulin and PLC 1 but not that between tubulin and G q. ␣ ␣ ␤ through a direct transfer of GTP from tubulin to G q. However, However, PIP2 did augment formation of tubulin–G q–PLC 1 ␤ ␤ at high tubulin concentrations, inhibition of PLC 1 is observed. complexes. Subsequent to potentiating PLC 1 activation, sus- ␤ This report demonstrates that tubulin inhibits PLC 1 by binding tained agonist-independent membrane binding of tubulin ␤ ␤ ␣ the PLC 1 substrate phosphatidylinositol 4,5-bisphosphate at PIP2- and PLC 1-rich sites appeared to inhibit G q coupling ␤ (PIP2 ). Tubulin binding of PIP2 was specific, because PIP2 but to PLC 1. Furthermore, colchicine increased membrane- ␤ not phosphatidylinositol 3,4,5-trisphosphate, phosphatidylino- associated tubulin and also inhibited PLC 1 activity in SK- sitol 3-phosphate, phosphatidylinositol, phosphatidylcholine, N-SH cells. Thus, tubulin, depending on local membrane con- phosphatidylethanolamine, or inositol 1,4,5-trisphosphate in- centration, may serve as a positive or negative regulator of hibited microtubule assembly.
    [Show full text]
  • IP3 Receptors – Lessons from Analyses Ex Cellula Ana M
    © 2018. Published by The Company of Biologists Ltd | Journal of Cell Science (2019) 132, jcs222463. doi:10.1242/jcs.222463 REVIEW SPECIAL ISSUE: RECONSTITUTING CELL BIOLOGY IP3 receptors – lessons from analyses ex cellula Ana M. Rossi and Colin W. Taylor* ABSTRACT and Ca2+ held within intracellular stores are entangled. For cardiac Inositol 1,4,5-trisphosphate receptors (IP Rs) are widely expressed muscle, depolarization of the plasma membrane (PM) causes 3 2+ intracellular channels that release Ca2+ from the endoplasmic voltage-gated Ca channels (Cav1.2, also known as CACNA1C) to 2+ reticulum (ER). We review how studies of IP Rs removed from their open, and the local increase in cytosolic free Ca concentration 3 2+ 2+ 2+ intracellular environment (‘ex cellula’), alongside similar analyses of ([Ca ]c) is then amplified by Ca -induced Ca release (CICR) ryanodine receptors, have contributed to understanding IP R through type 2 ryanodine receptors (RyR2) in the sarcoplasmic 3 2+ behaviour. Analyses of permeabilized cells have demonstrated that reticulum (Bers, 2002) (Fig. 1A). CICR and the local Ca 2+ signalling that is required to avoid CICR from becoming the ER is the major intracellular Ca store, and that IP3 stimulates Ca2+ release from this store. Radioligand binding confirmed that the explosive have become recurrent themes in the field of Ca2+ signalling (Rios, 2018). Fluorescent Ca2+ indicators and 4,5-phosphates of IP3 are essential for activating IP3Rs, and optical microscopy now allow Ca2+ sparks, local Ca2+ signals facilitated IP3R purification and cloning, which paved the way for evoked by a small cluster of RyRs, to be measured with exquisite structural analyses.
    [Show full text]
  • Rapid Stimulation of Cellular Pi Uptake by the Inositol Pyrophosphate Insp8
    Chemical Science View Article Online EDGE ARTICLE View Journal | View Issue Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP induced by its Cite this: Chem. Sci., 2020, 11,10265 8 All publication charges for this article photothermal release from lipid nanocarriers using have been paid for by the Royal Society † of Chemistry a near infra-red light-emitting diode Zhenzhen Wang,‡a Nikolaus Jork, b Tamara Bittner, b Huanchen Wang, a Henning J. Jessen b and Stephen B. Shears *a Inositol pyrophosphates (PP-InsPs), including diphospho-myo-inositol pentakisphosphate (5-InsP7) and bis-diphospho-myo-inositol tetrakisphosphate (1,5-InsP8), are highly polar, membrane-impermeant signaling molecules that control many homeostatic responses to metabolic and bioenergetic imbalance. To delineate their molecular activities, there is an increasing need for a toolbox of methodologies for real-time modulation of PP-InsP levels inside large populations of cultured cells. Here, we describe procedures to package PP-InsPs into thermosensitive phospholipid nanocapsules that are impregnated Creative Commons Attribution 3.0 Unported Licence. with a near infra-red photothermal dye; these liposomes are readily accumulated into cultured cells. The PP-InsPs remain trapped inside the liposomes until the cultures are illuminated with a near infra-red light-emitting diode (LED) which permeabilizes the liposomes to promote PP-InsP release. Additionally, so as to optimize these procedures, a novel stably fluorescent 5-InsP7 analogue (i.e., 5-FAM-InsP7) was synthesized with the assistance of click-chemistry; the delivery and deposition of the analogue inside cells was monitored by flow cytometry and by confocal microscopy.
    [Show full text]
  • Role of Inositols and Inositol Phosphates in Energy Metabolism
    molecules Review Role of Inositols and Inositol Phosphates in Energy Metabolism Saimai Chatree 1, Nanthaphop Thongmaen 2, Kwanchanit Tantivejkul 3, Chantacha Sitticharoon 2 and Ivana Vucenik 4,5,* 1 Faculty of Medicine and Public Health, HRH Princess Chulabhorn College of Medical Science, Chulabhorn Royal Academy, Bangkok 10210, Thailand; [email protected] 2 Department of Physiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; [email protected] (N.T.); [email protected] (C.S.) 3 Sugavia Co., Ltd., Nakhonratchasima 30130, Thailand; [email protected] 4 Department of Medical and Research Technology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA 5 Department of Pathology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA * Correspondence: [email protected]; Tel.: +1-410-706-1832; Fax: +1-410-706-5229 Academic Editor: Stephen Shears Received: 3 October 2020; Accepted: 27 October 2020; Published: 1 November 2020 Abstract: Recently, inositols, especially myo-inositol and inositol hexakisphosphate, also known as phytic acid or IP6, with their biological activities received much attention for their role in multiple health beneficial effects. Although their roles in cancer treatment and prevention have been extensively reported, interestingly, they may also have distinctive properties in energy metabolism and metabolic disorders. We review inositols and inositol phosphate metabolism in mammalian cells to establish their biological activities and highlight their potential roles in energy metabolism. These molecules are known to decrease insulin resistance, increase insulin sensitivity, and have diverse properties with importance from cell signaling to metabolism. Evidence showed that inositol phosphates might enhance the browning of white adipocytes and directly improve insulin sensitivity through adipocytes.
    [Show full text]
  • Lnositol 1,4,5=Trisphosphate Receptor Binding: Autoradiographic Localization in Rat Brain
    The Journal of Neuroscience, January 1989, 9(l): 339-348 lnositol 1,4,5=Trisphosphate Receptor Binding: Autoradiographic Localization in Rat Brain Paul F. Worley,1s2 Jay M. Baraban,lv3 and Solomon H. Snyderi.3,4 Departments of ‘Neuroscience, *Neurology, 3Psychiatry and Behavioral Sciences, and 4Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 lnositol 1,4,5trisphosphate is a second messenger gener- al., 1987a). In the present study, we have employed autora- ated by stimulation of the phosphoinositide cycle, thought diography to localize IP, receptors in brain in greater detail, to release calcium from intracellular stores. We have mapped comparing their distribution with that of PKC and adenylate the distribution of 3H-inositol 1,4,5trisphosphate receptor cyclase, labeled by 3H-phorbol 12,13-dibutyrate (3H-PDBu) and binding sites in rat brain by autoradiographic techniques. The 3H-forskolin, respectively. We have also characterized the on- cerebellum contains the highest level of inositol 1,4,5tris- togeny of IP, receptor binding in brain and performed lesion phosphate binding sites in brain, which appear to be selec- studies to identify neuronal elements enriched in IP, receptors. tively localized to Purkinje cells. Moderate levels of binding sites are present in the hippocampus, cerebral cortex, cau- Materials and Methods date, and substantia nigra. Lesion studies indicate that bind- *H-IP, autoradiography.3H-IP, autoradiography was carried out as de- ing in the hippocampus is restricted to intrinsic neuronal scribed (Worley et al., 1987a). Male Sprague-Dawley rats were anes- thetized with pentobarbital and perfused via the left cardiac ventricle elements and in the nigra is found on terminals of the stria- with buffer containing 50 mM sodium phosphate, pH 7.5, and 100 mM tonigral projection.
    [Show full text]
  • Biochemical and Functional Responses Stimulated by Platelet-Activating Factor in Murine Peritoneal Macrophages
    Biochemical and Functional Responses Stimulated by Platelet-activating Factor in Murine Peritoneal Macrophages Veronica Prpic,* Ronald J. Uhing,* James E. Weiel,* Lazlo Jakoi,~ Govind Gawdi,§ Brian Herman,§ and Dolph O. Adams*ll Departments of * Pathology, II Microbiology-Immunology, and eMedicine, Duke University, Durham, North Carolina 27710; and the §Department of Anatomy, University of North Carolina, Chapel Hill, North Carolina 27514 Abstract. Platelet-activating factor (PAF) is a potent the macrophages. PAF also led to increases of 1,2- stimulant of leukocytes, including macrophages. To diacylglycerol of • 200 pmol/107 cells. A characteristic analyze the mechanisms of its effects upon macro- pattern of enhanced protein phosphorylation, similar to Downloaded from http://rupress.org/jcb/article-pdf/107/1/363/1056534/363.pdf by guest on 02 February 2021 phages, we determined whether macrophages bear that initiated by both phorbol 12,13-myristate and lipo- specific surface receptors for PAE By competitive ra- polysaccharide, was observed and involved enhanced dioactive binding assays, we determined two classes of phosphorylation of proteins of 28, 33, 67, and 103 kD. specific receptors to be present on purified membranes The half-maximal dose of PAF for initiating all the derived from murine peritoneal macrophages (one hav- above effects was ,,,,5 x 10-9 M. PAF also initiated ing a Kd of '~1 X 10 -'° M and one a Ka of ~ 2 x 10-9 significant chemotaxis of the cells; the half-maximal M). When the macrophages were incubated with PAF, dose for this effect was ~1 x 10-" M. Taken together, rapid formation of several inositol phosphates includ- these observations suggest that murine mononuclear ing inositol 1,4,5-trisphosphate and inositol 1,3,4,5- phagocytes bear specific membrane receptors for PAF tetrakisphosphate were observed.
    [Show full text]