DOCSLIB.ORG

3-Hydroxyglutaric Acid Is Transported Via the Sodium-Dependent Dicarboxylate Transporter Nadc3

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
3-Hydroxyglutaric Acid Is Transported Via the Sodium-Dependent Dicarboxylate Transporter Nadc3 J Mol Med (2007) 85:763–770 DOI 10.1007/s00109-007-0174-5 RAPID COMMUNICATION 3-Hydroxyglutaric acid is transported via the sodium-dependent dicarboxylate transporter NaDC3 Franziska Stellmer & Britta Keyser & Birgitta C. Burckhardt & Hermann Koepsell & Thomas Streichert & Markus Glatzel & Sabrina Jabs & Joachim Thiem & Wilhelm Herdering & David M. Koeller & Stephen I. Goodman & Zoltan Lukacs & Kurt Ullrich & Gerhard Burckhardt & Thomas Braulke & Chris Mühlhausen Received: 20 October 2006 /Revised: 16 January 2007 /Accepted: 8 February 2007 / Published online: 14 March 2007 # Springer-Verlag 2007 Abstract Patients with glutaryl-CoA dehydrogenase (GCDH) deficiency accumulate glutaric acid (GA) and 3- hydroxyglutaric acid (3OH-GA) in their blood and urine. To identify the transporter mediating the translocation of 3OH- Electronic supplementary material The online version of this article (doi:10.1007/s00109-007-0174-5) contains supplementary material, which is available to authorized users. F. Stellmer : B. Keyser : S. Jabs : Z. Lukacs : K. Ullrich : T. Braulke : C. Mühlhausen (*) Department of Pediatrics, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany FRANZISKA STELLMER CHRIS MÜHLHAUSEN e-mail: [email protected] just completed her MD thesis in received his MD from the Georg- the Molecular Biology Lab of August-University of Göttingen B. C. Burckhardt : G. Burckhardt the Department of Pediatrics, after having performed a thesis Center of Physiology and Pathophysiology, University Medical Center about transport of lysosomal Georg-August-University, Hamburg-Eppendorf. She will proteins in the Department of Humboldtallee 23, qualify as a physician shortly. Biochemistry with Prof. K. von 37073 Göttingen, Germany Her research projects include Figura. He is presently a spe- renal abnormalities in a mouse cialist pediatrician and research H. Koepsell model of glutaryl-CoA dehy- project leader at the Department Institute of Anatomy and Cell Biology, University of Würzburg, drogenase deficiency, especial- of Pediatrics, University Medical Koellikerstrasse 6, ly renal transport mechanisms. Center Hamburg-Eppendorf. His 97070 Würzburg, Germany research interests include inborn metabolic diseases, especially T. Streichert : Z. Lukacs glutaryl-CoA dehydrogenase de- Department of Clinical Chemistry, University Medical Center ficiency as well as lysosomal Hamburg-Eppendorf, storage diseases. Martinistrasse 52, 20246 Hamburg, Germany GA through membranes, kidney tissue of Gcdh−/− mice have M. Glatzel been investigated because of its central role in urinary Department of Neuropathology, excretion of this metabolite. Using microarray analyses of University Medical Center Hamburg-Eppendorf, − − Martinistrasse 52, kidney-expressed genes in Gcdh / mice, several differen- 20246 Hamburg, Germany tially expressed genes encoding transporter proteins were 764 J Mol Med (2007) 85:763–770 identified. Real-time polymerase chain reaction analysis tissues and body fluids is accompanied by irreversible confirmed the upregulation of the sodium-dependent dicar- destruction of striatal neurons with subsequent development boxylate cotransporter 3 (NaDC3) and the organic cation of an irreversible disabling movement disorder [1]. transporter 2 (OCT2). Expression analysis of NaDC3 in Whereas the pathomechanisms in GA1 causing striatal Xenopus laevis oocytes by the two-electrode-voltage-clamp dysfunction and degeneration are not fully understood, both technique demonstrated the sodium-dependent translocation N-methyl-D-aspartate receptor-mediated neurotoxicity and of 3OH-GA with a KM value of 0.95 mM. Furthermore, disintegration of endothelial barriers by 3OH-GA are tracer flux measurements in Chinese hamster ovary cells discussed [2–5]. However, cellular mechanisms facilitating overexpressing OCT2 showed that 3OH-GA inhibited the transport of 3OH-GA through membranes have not significantly the uptake of methyl-4-phenylpyridinium, been investigated yet. whereas 3OH-GA is not transported by OCT2. The data A mouse model of GA1 was generated by a targeted demonstrate for the first time the membrane translocation of disruption of the Gcdh gene with a biochemical phenotype 3OH-GA mediated by NaDC3 and the cis-inhibitory effect similar to human GA1 patients, including high excretion on OCT2-mediated transport of cations. rates of GA and 3OH-GA into the urine [6]. Additionally, diffuse spongiform myelinopathy and an unexplained Keywords Glutaric aciduria type 1 . Slc22a2 . OCT2 . hypertrophy of kidneys were seen in Gcdh−/− mice. Glutaryl-CoA dehydrogenase deficiency. Slc13a3 . NaDC3 To examine transporters involved in the translocation of 3OH-GA via membranes, we performed oligonucleotide Abbreviations microarray analyses in kidneys of Gcdh-deficient mice and GA1 glutaric aciduria type 1 found several solute carrier protein (Slc) genes differential- GA glutaric acid ly regulated. Confirmed by quantitative real-time polymer- Gcdh glutaryl-CoA dehydrogenase ase chain reaction (qRT-PCR), immunohistochemistry, NaDC3 sodium-dependent dicarboxylate expression analyses in Xenopus laevis oocytes and two- cotransporter 3 electrode-voltage-clamp measurements, the sodium-depen- 3OH- 3-hydroxyglutaric acid dent dicarboxylate cotransporter 3 (NaDC3, Slc13a3) was GA identified as a potential transporter mediating the low OCT2 organic cation transporter 2 affinity translocation of 3OH-GA at the basolateral mem- Slc solute carrier brane of kidney proximal tubule cells. Materials and methods Introduction Materials 1-[3H]-Methyl-4-phenylpyridinium (MPP) and Glutaric aciduria type 1 (GA1) is caused by the deficiency [3H]-sodium borohydride (NaBH ) were purchased from of the mitochondrial matrix protein glutaryl-CoA dehydro- 4 Biotrend (Köln, Germany) and Amersham Pharmacia genase (GCDH). The enzyme catalyzes the oxidative (Freiburg, Germany) respectively. Dimethyl-3-ketoglutaric decarboxylation of glutaryl-CoA to crotonyl-CoA in the and GA acids and diaminobenzidine were obtained from catabolic pathway of lysine, hydroxylysine, and tryptophan. Sigma Chemical (St. Louis, MO). Silica gel plates (HPTLC During catabolic crises, the accumulation of large amounts F254) and silica gel (0.015–0.063 mesh) were from Merck of glutaric (GA) and 3-hydroxyglutaric (3OH-GA) acids in (Darmstadt, Germany). 3OH-GA was synthesized as 3 : reported previously [4]. The synthesis of [ H]-labeled J. Thiem W. Herdering 3OH-GA is described in the supplementary methods online. Institute of Organic Chemistry, University of Hamburg, The human (h) NaDC3 complementary deoxyribonucleic Martin-Luther-King-Platz 6, 20146 Hamburg, Germany acid (cDNA) was kindly provided by Dr. V. Ganapathy (Medical College of Georgia, Augusta, GA). All other D. M. Koeller chemicals were of analytical grade or higher. Department of Pediatrics, Oregon Health and Science University, 707 SW Gaines Road, Portland, OR 97239, USA Antibodies Polyclonal anti-OCT2 and anti-NaDC3 anti- bodies were from Alpha Diagnostic International (San S. I. Goodman Antonio, TX), anti-actin from Santa Cruz (Heidelberg, Department of Pediatrics, Germany), and horseradish peroxidase (HRP)- and alkaline- University of Colorado Health Sciences Center, P.O. Box C233, 4200 East Ninth Avenue, phosphatase-conjugated anti-rabbit and anti-guinea-pig Denver, CO 80262, USA immunoglobulin G (IgG), respectively, from Jackson J Mol Med (2007) 85:763–770 765 Immunoresearch (West Grove, PA). The polyclonal guinea- cRNA synthesis and oocyte injection Stage V–VI oocytes pig antibody against the basolateral K+/Cl− (K-Cl) cotrans- from X. laevis (Nasco, Fort Atkinson, WI) were defollicu- porter Kcc4 (Slc12a7) [7] was kindly provided by Dr. C. lated by an overnight incubation at 18°C with collagenase Hübner (University Medical Center Hamburg-Eppendorf). (Type CLSII, Biochrom KG, Germany; 0.5 mg/ml) in oocyte Ringer’s solution (ORI) containing 110 mM NaCl, Mice-Gcdh −/− mice and wild-type littermate controls were 3 mM KCl, 2 mM CaCl2, 5 mM HEPES/Tris, pH 7.5. One generated from heterozygotes [6]. The genetic background day after removal from the ovaries, oocytes were injected of all mice groups used in this study was C57BL6/SJ129 with 23 nl of water or 23 ng hNaDC3-cRNA in an hybrid. The genotypes were confirmed by polymerase chain equivalent volume, synthesized from NotI-linearized plas- reaction (PCR) and measurements of glutarylcarnitine mid (mMessage mMachine-T7 in vitro transcription kit; concentration in dried blood spots. The mice were housed Ambion, Austin, TX). After injection, the oocytes were in an animal facility of the University Hospital with a incubated for 3 days at 18°C in ORI, supplemented with 12-h light–dark cycle and allowed water and food ad 50 μg/ml gentamycin and 2.5 mM sodium pyruvate. libitum. Animal care was provided in accordance with institutional guidelines. Anesthesized mice were either used Electrophysiological studies Current recordings were per- for preparation of kidneys or perfused with phosphate- formed in ORI using the two-electrode-voltage-clamp buffered saline, pH 7.4, containing 50 U/100 ml heparin technique with a commercial amplifier (OC725, Warner, followed by cryosectioning. Hambden, CT). Borosilicate glass microelectrodes were filled with 3 M KCl and had resistances of ∼1MΩ. The Target labeling and microarray hybridization Procedures resting membrane potential of the oocytes ranged between for cDNA synthesis, labeling, and hybridization were −22 and −36 mV, and the holding currents to achieve
Load more

Recommended publications
  • Screening and Identification of Key Biomarkers in Clear Cell Renal Cell Carcinoma Based on Bioinformatics Analysis
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.21.423889; this version posted December 23, 2020. 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. Screening and identification of key biomarkers in clear cell renal cell carcinoma based on bioinformatics analysis Basavaraj Vastrad1, Chanabasayya Vastrad*2 , Iranna Kotturshetti 1. Department of Biochemistry, Basaveshwar College of Pharmacy, Gadag, Karnataka 582103, India. 2. Biostatistics and Bioinformatics, Chanabasava Nilaya, Bharthinagar, Dharwad 580001, Karanataka, India. 3. Department of Ayurveda, Rajiv Gandhi Education Society`s Ayurvedic Medical College, Ron, Karnataka 562209, India. * Chanabasayya Vastrad [email protected] Ph: +919480073398 Chanabasava Nilaya, Bharthinagar, Dharwad 580001 , Karanataka, India bioRxiv preprint doi: https://doi.org/10.1101/2020.12.21.423889; this version posted December 23, 2020. 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 Clear cell renal cell carcinoma (ccRCC) is one of the most common types of malignancy of the urinary system. The pathogenesis and effective diagnosis of ccRCC have become popular topics for research in the previous decade. In the current study, an integrated bioinformatics analysis was performed to identify core genes associated in ccRCC. An expression dataset (GSE105261) was downloaded from the Gene Expression Omnibus database, and included 26 ccRCC and 9 normal kideny samples. Assessment of the microarray dataset led to the recognition of differentially expressed genes (DEGs), which was subsequently used for pathway and gene ontology (GO) enrichment analysis.
    [Show full text]
  • New Advances in Urea Transporter UT-A1 Membrane Trafficking
    Int. J. Mol. Sci. 2013, 14, 10674-10682; doi:10.3390/ijms140510674 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review New Advances in Urea Transporter UT-A1 Membrane Trafficking Guangping Chen Department of Physiology, Emory University School of Medicine, Atlanta, GA 30322, USA; E-Mail: [email protected]; Tel.: +1-404-727-7494; Fax: +1-404-727-2648. Received: 22 April 2013; in revised form: 9 May 2013 / Accepted: 9 May 2013 / Published: 21 May 2013 Abstract: The vasopressin-regulated urea transporter UT-A1, expressed in kidney inner medullary collecting duct (IMCD) epithelial cells, plays a critical role in the urinary concentrating mechanisms. As a membrane protein, the function of UT-A1 transport activity relies on its presence in the plasma membrane. Therefore, UT-A1 successfully trafficking to the apical membrane of the polarized epithelial cells is crucial for the regulation of urea transport. This review summarizes the research progress of UT-A1 regulation over the past few years, specifically on the regulation of UT-A1 membrane trafficking by lipid rafts, N-linked glycosylation and a group of accessory proteins. Keywords: lipid rafts; glycosylation; accessory proteins; SNARE protein; cytoskeleton protein 1. Introduction Urea is the major end product of amino acid metabolism. It is generated from the ornithine cycle in liver, and is ultimately excreted by the kidney representing 90% of total nitrogen in urine. The physiological significance of urea in the production of concentrated urine was recognized by Gamble in the 1930s [1,2]. Urea reabsorbed in the kidney inner medullary collecting duct (IMCD) contributes to the development of the osmolality in the medullary interstitium.
    [Show full text]
  • Small Cell Ovarian Carcinoma: Genomic Stability and Responsiveness to Therapeutics
    Gamwell et al. Orphanet Journal of Rare Diseases 2013, 8:33 http://www.ojrd.com/content/8/1/33 RESEARCH Open Access Small cell ovarian carcinoma: genomic stability and responsiveness to therapeutics Lisa F Gamwell1,2, Karen Gambaro3, Maria Merziotis2, Colleen Crane2, Suzanna L Arcand4, Valerie Bourada1,2, Christopher Davis2, Jeremy A Squire6, David G Huntsman7,8, Patricia N Tonin3,4,5 and Barbara C Vanderhyden1,2* Abstract Background: The biology of small cell ovarian carcinoma of the hypercalcemic type (SCCOHT), which is a rare and aggressive form of ovarian cancer, is poorly understood. Tumourigenicity, in vitro growth characteristics, genetic and genomic anomalies, and sensitivity to standard and novel chemotherapeutic treatments were investigated in the unique SCCOHT cell line, BIN-67, to provide further insight in the biology of this rare type of ovarian cancer. Method: The tumourigenic potential of BIN-67 cells was determined and the tumours formed in a xenograft model was compared to human SCCOHT. DNA sequencing, spectral karyotyping and high density SNP array analysis was performed. The sensitivity of the BIN-67 cells to standard chemotherapeutic agents and to vesicular stomatitis virus (VSV) and the JX-594 vaccinia virus was tested. Results: BIN-67 cells were capable of forming spheroids in hanging drop cultures. When xenografted into immunodeficient mice, BIN-67 cells developed into tumours that reflected the hypercalcemia and histology of human SCCOHT, notably intense expression of WT-1 and vimentin, and lack of expression of inhibin. Somatic mutations in TP53 and the most common activating mutations in KRAS and BRAF were not found in BIN-67 cells by DNA sequencing.
    [Show full text]
  • Viewed Under 23 (B) Or 203 (C) fi M M Male Cko Mice, and Largely Unaffected Magni Cation; Scale Bars, 500 M (B) and 50 M (C)
    BRIEF COMMUNICATION www.jasn.org Renal Fanconi Syndrome and Hypophosphatemic Rickets in the Absence of Xenotropic and Polytropic Retroviral Receptor in the Nephron Camille Ansermet,* Matthias B. Moor,* Gabriel Centeno,* Muriel Auberson,* † † ‡ Dorothy Zhang Hu, Roland Baron, Svetlana Nikolaeva,* Barbara Haenzi,* | Natalya Katanaeva,* Ivan Gautschi,* Vladimir Katanaev,*§ Samuel Rotman, Robert Koesters,¶ †† Laurent Schild,* Sylvain Pradervand,** Olivier Bonny,* and Dmitri Firsov* BRIEF COMMUNICATION *Department of Pharmacology and Toxicology and **Genomic Technologies Facility, University of Lausanne, Lausanne, Switzerland; †Department of Oral Medicine, Infection, and Immunity, Harvard School of Dental Medicine, Boston, Massachusetts; ‡Institute of Evolutionary Physiology and Biochemistry, St. Petersburg, Russia; §School of Biomedicine, Far Eastern Federal University, Vladivostok, Russia; |Services of Pathology and ††Nephrology, Department of Medicine, University Hospital of Lausanne, Lausanne, Switzerland; and ¶Université Pierre et Marie Curie, Paris, France ABSTRACT Tight control of extracellular and intracellular inorganic phosphate (Pi) levels is crit- leaves.4 Most recently, Legati et al. have ical to most biochemical and physiologic processes. Urinary Pi is freely filtered at the shown an association between genetic kidney glomerulus and is reabsorbed in the renal tubule by the action of the apical polymorphisms in Xpr1 and primary fa- sodium-dependent phosphate transporters, NaPi-IIa/NaPi-IIc/Pit2. However, the milial brain calcification disorder.5 How- molecular identity of the protein(s) participating in the basolateral Pi efflux remains ever, the role of XPR1 in the maintenance unknown. Evidence has suggested that xenotropic and polytropic retroviral recep- of Pi homeostasis remains unknown. Here, tor 1 (XPR1) might be involved in this process. Here, we show that conditional in- we addressed this issue in mice deficient for activation of Xpr1 in the renal tubule in mice resulted in impaired renal Pi Xpr1 in the nephron.
    [Show full text]
  • Differential Expression of Hydroxyurea Transporters in Normal and Polycythemia Vera Hematopoietic Stem and Progenitor Cell Subpopulations
    Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2021 Differential expression of hydroxyurea transporters in normal and polycythemia vera hematopoietic stem and progenitor cell subpopulations Tan, Ge ; Meier-Abt, Fabienne Abstract: Polycythemia vera (PV) is a myeloproliferative neoplasm marked by hyperproliferation of the myeloid lineages and the presence of an activating JAK2 mutation. Hydroxyurea (HU) is a standard treat- ment for high-risk patients with PV. Because disease-driving mechanisms are thought to arise in PV stem cells, effective treatments should target primarily the stem cell compartment. We tested for theantipro- liferative effect of patient treatment with HU in fluorescence-activated cell sorting-isolated hematopoietic stem/multipotent progenitor cells (HSC/MPPs) and more committed erythroid progenitors (common myeloid/megakaryocyte-erythrocyte progenitors [CMP/MEPs]) in PV using RNA-sequencing and gene set enrichment analysis. HU treatment led to significant downregulation of gene sets associated with cell proliferation in PV HSCs/MPPs, but not in PV CMP/MEPs. To explore the mechanism underlying this finding, we assessed for expression of solute carrier membrane transporters, which mediate trans- membrane movement of drugs such as HU into target cells. The active HU uptake transporter OCTN1 was upregulated in HSC/MPPs compared with CMP/MEPs of untreated patients with PV, and the HU diffusion facilitator urea transporter B (UTB) was downregulated in HSC/MPPs compared withCM- P/MEPs in all patient and control groups tested. These findings indicate a higher accumulation ofHU within PV HSC/MPPs compared with PV CMP/MEPs and provide an explanation for the differential effects of HU in HSC/MPPs and CMP/MEPs of patients with PV.
    [Show full text]
  • The Concise Guide to Pharmacology 2019/20
    Edinburgh Research Explorer THE CONCISE GUIDE TO PHARMACOLOGY 2019/20 Citation for published version: Cgtp Collaborators 2019, 'THE CONCISE GUIDE TO PHARMACOLOGY 2019/20: Transporters', British Journal of Pharmacology, vol. 176 Suppl 1, pp. S397-S493. https://doi.org/10.1111/bph.14753 Digital Object Identifier (DOI): 10.1111/bph.14753 Link: Link to publication record in Edinburgh Research Explorer Document Version: Publisher's PDF, also known as Version of record Published In: British Journal of Pharmacology General rights Copyright for the publications made accessible via the Edinburgh Research Explorer is retained by the author(s) and / or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights. Take down policy The University of Edinburgh has made every reasonable effort to ensure that Edinburgh Research Explorer content complies with UK legislation. If you believe that the public display of this file breaches copyright please contact [email protected] providing details, and we will remove access to the work immediately and investigate your claim. Download date: 28. Sep. 2021 S.P.H. Alexander et al. The Concise Guide to PHARMACOLOGY 2019/20: Transporters. British Journal of Pharmacology (2019) 176, S397–S493 THE CONCISE GUIDE TO PHARMACOLOGY 2019/20: Transporters Stephen PH Alexander1 , Eamonn Kelly2, Alistair Mathie3 ,JohnAPeters4 , Emma L Veale3 , Jane F Armstrong5 , Elena Faccenda5 ,SimonDHarding5 ,AdamJPawson5 , Joanna L
    [Show full text]
  • Dur3 Is the Major Urea Transporter in Candida Albicans and Is Co-Regulated with the Urea Amidolyase Dur1,2 Dhammika H
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Kenneth Nickerson Papers Papers in the Biological Sciences 2011 Dur3 is the major urea transporter in Candida albicans and is co-regulated with the urea amidolyase Dur1,2 Dhammika H. M. L. P Navarathna National Cancer Institute Aditi Das Universitat Wurzburg Joachim Morschhauser Universitat Wurzburg Kenneth W. Nickerson University of Nebraska - Lincoln, [email protected] David D. Roberts National Cancer Institute, [email protected] Follow this and additional works at: http://digitalcommons.unl.edu/bioscinickerson Part of the Environmental Microbiology and Microbial Ecology Commons, Other Life Sciences Commons, and the Pathogenic Microbiology Commons Navarathna, Dhammika H. M. L. P; Das, Aditi; Morschhauser, Joachim; Nickerson, Kenneth W.; and Roberts, David D., "Dur3 is the major urea transporter in Candida albicans and is co-regulated with the urea amidolyase Dur1,2" (2011). Kenneth Nickerson Papers. 11. http://digitalcommons.unl.edu/bioscinickerson/11 This Article is brought to you for free and open access by the Papers in the Biological Sciences at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Kenneth Nickerson Papers by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Microbiology (2011), 157, 270–279 DOI 10.1099/mic.0.045005-0 Dur3 is the major urea transporter in Candida albicans and is co-regulated with the urea amidolyase Dur1,2 Dhammika H. M. L. P. Navarathna,1 Aditi Das,2 Joachim Morschha¨user,2 Kenneth W. Nickerson3 and David D. Roberts1 Correspondence 1Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes David D.
    [Show full text]
  • Abrothrix Olivacea)
    Received: 13 June 2017 | Revised: 30 May 2018 | Accepted: 31 May 2018 DOI: 10.1111/mec.14778 ORIGINAL ARTICLE An association between differential expression and genetic divergence in the Patagonian olive mouse (Abrothrix olivacea) Facundo M. Giorello1,2 | Matias Feijoo1 | Guillermo D'Elía3 | Daniel E. Naya1 | Lourdes Valdez3 | Juan C. Opazo3 | Enrique P. Lessa1 1Departamento de Ecología y Evolución, Facultad de Ciencias, Universidad de la Abstract República, Montevideo, Uruguay Recent molecular studies have found striking differences between desert‐adapted 2Espacio de Biología Vegetal del Noreste, species and model mammals regarding water conservation. In particular, aquaporin Centro Universitario de Tacuarembó, Universidad de la República, Tacuarembó, 4, a classical gene involved in water regulation of model species, is absent or not Uruguay expressed in the kidneys of desert‐adapted species. To further understand the 3Instituto de Ciencias Ambientales y Evolutivas, Universidad Austral de Chile, molecular response to water availability, we studied the Patagonian olive mouse Valdivia, Chile Abrothrix olivacea, a species with an unusually broad ecological tolerance that exhi- Correspondence bits a great urine concentration capability. The species is able to occupy both the Facundo M. Giorello, Departamento de arid Patagonian steppe and the Valdivian and Magellanic forests. We sampled 95 Ecología y Evolución, Facultad de Ciencias, Universidad de la República, Iguá 4225, olive mouse specimens from four localities (two in the steppe and two in the for- Montevideo 11400, Uruguay. ests) and analysed both phenotypic variables and transcriptomic data to investigate Email: [email protected] the response of this species to the contrasting environmental conditions. The rela- Funding information tive size of the kidney and the ratio of urine to plasma concentrations were, as Fondo Nacional de Desarrollo Científico y Tecnológico, Grant/Award Number: expected, negatively correlated with annual rainfall.
    [Show full text]
  • Structure and Permeation Mechanism of a Mammalian Urea Transporter
    Structure and permeation mechanism of a mammalian urea transporter Elena J. Levina,1, Yu Caoa,1, Giray Enkavib, Matthias Quickc, Yaping Pana, Emad Tajkhorshidb,2, and Ming Zhoua,2 aDepartment of Physiology and Cellular Biophysics, College of Physicians and Surgeons, Columbia University, 630 West 168th Street, New York, NY 10032; bCenter for Biophysics and Computational Biology, Department of Biochemistry, College of Medicine, and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801; and cDepartment of Psychiatry and Center for Molecular Recognition, Columbia University, 650 West 168th Street, New York, NY 10032 Edited by Christopher Miller, HHMI, Brandeis University, Waltham, MA, and approved June 1, 2012 (received for review May 3, 2012) As an adaptation to infrequent access to water, terrestrial mam- homolog (21), dvUT, which forms a trimer with a continuous mals produce urine that is hyperosmotic to plasma. To prevent membrane-spanning pore at the center of each protomer. How- osmotic diuresis by the large quantity of urea generated by protein ever, it remained unclear how similar this structure was to that catabolism, the kidney epithelia contain facilitative urea transpor- of the mammalian UTs, and the details of the permeation me- ters (UTs) that allow rapid equilibration between the urinary space chanism were unknown. To answer these questions, we solved and the hyperosmotic interstitium. Here we report the first X-ray the structure of a mammalian UT-B and investigated the permea- crystal structure of a mammalian UT, UT-B, at a resolution of 2.36 Å. tion mechanism with molecular dynamics simulations and func- UT-B is a homotrimer and each protomer contains a urea con- tional studies of UT-B mutants.
    [Show full text]
  • Dur3 Is the Major Urea Transporter in Candida Albicans and Is Co-Regulated with the Urea Amidolyase Dur1,2
    CORE Metadata, citation and similar papers at core.ac.uk Provided by PubMed Central Microbiology (2011), 157, 270–279 DOI 10.1099/mic.0.045005-0 Dur3 is the major urea transporter in Candida albicans and is co-regulated with the urea amidolyase Dur1,2 Dhammika H. M. L. P. Navarathna,1 Aditi Das,2 Joachim Morschha¨user,2 Kenneth W. Nickerson3 and David D. Roberts1 Correspondence 1Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes David D. Roberts of Health, Bethesda, MD 20892-1500, USA [email protected] 2Institut fu¨r Molekulare Infektionsbiologie, Universita¨t Wu¨rzburg, Wu¨rzburg, Germany 3School of Biological Sciences, University of Nebraska, Lincoln, NE, USA Hemiascomycetes, including the pathogen Candida albicans, acquire nitrogen from urea using the urea amidolyase Dur1,2, whereas all other higher fungi use primarily the nickel-containing urease. Urea metabolism via Dur1,2 is important for resistance to innate host immunity in C. albicans infections. To further characterize urea metabolism in C. albicans we examined the function of seven putative urea transporters. Gene disruption established that Dur3, encoded by orf 19.781, is the predominant transporter. [14C]Urea uptake was energy-dependent and decreased approximately sevenfold in a dur3D mutant. DUR1,2 and DUR3 expression was strongly induced by urea, whereas the other putative transporter genes were induced less than twofold. Immediate induction of DUR3 by urea was independent of its metabolism via Dur1,2, but further slow induction of DUR3 required the Dur1,2 pathway. We investigated the role of the GATA transcription factors Gat1 and Gln3 in DUR1,2 and DUR3 expression.
    [Show full text]
  • Tricarboxylic Acid Cycle Intermediates As Myometabokines
    H OH metabolites OH Review Signals from the Circle: Tricarboxylic Acid Cycle Intermediates as Myometabokines Jennifer Maurer 1 , Miriam Hoene 1 and Cora Weigert 1,2,3,* 1 Department for Diagnostic Laboratory Medicine, Institute for Clinical Chemistry and Pathobiochemistry, University Hospital Tuebingen, 72076 Tuebingen, Germany; [email protected] (J.M.); [email protected] (M.H.) 2 Institute for Diabetes Research and Metabolic Diseases, Helmholtz Center Munich, University of Tuebingen, 72076 Tuebingen, Germany 3 German Center for Diabetes Research (DZD), 85764 Oberschleissheim, Germany * Correspondence: [email protected]; Tel.: +49-7071-29-85670 Abstract: Regular physical activity is an effective strategy to prevent and ameliorate aging-associated diseases. In particular, training increases muscle performance and improves whole-body metabolism. Since exercise affects the whole organism, it has countless health benefits. The systemic effects of exercise can, in part, be explained by communication between the contracting skeletal muscle and other organs and cell types. While small proteins and peptides known as myokines are the most prominent candidates to mediate this tissue cross-talk, recent investigations have paid increasing attention to metabolites. The purpose of this review is to highlight the potential role of tricarboxylic acid (TCA) metabolites as humoral mediators of exercise adaptation processes. We focus on TCA metabolites that are released from human skeletal muscle in response to exercise and provide an overview of their potential auto-, para- or endocrine health-promoting effects. Keywords: TCA cycle; exercise; myometabokine; exercise adaptation; liver; arterio-venous difference; Citation: Maurer, J.; Hoene, M.; Weigert, C. Signals from the Circle: succinate; citrate Tricarboxylic Acid Cycle Intermediates as Myometabokines.
    [Show full text]
  • H00008170-Q01 規格 : [ 10 Ug ] [ 25 Ug ] List All
    SLC14A2 (Human) Recombinant Protein (Q01) Catalog # : H00008170-Q01 規格 : [ 10 ug ] [ 25 ug ] List All Specification Application Image Product Human SLC14A2 partial ORF ( NP_009094, 40 a.a. - 128 a.a.) Enzyme-linked Immunoabsorbent Assay Description: recombinant protein with GST-tag at N-terminal. Western Blot (Recombinant Sequence: ALPLLEMPEEKDLRSSNEDSHIVKIEKLNERSKRKDDGVAHRDSAGQRCI protein) CLSKAVGYLTGDMKEYRIWLKDKHLALQFIDWVLRGTAQ Antibody Production Host: Wheat Germ (in vitro) Protein Array Theoretical MW 35.53 (kDa): Preparation in vitro wheat germ expression system Method: Purification: Glutathione Sepharose 4 Fast Flow Quality Control 12.5% SDS-PAGE Stained with Coomassie Blue. Testing: Storage Buffer: 50 mM Tris-HCI, 10 mM reduced Glutathione, pH=8.0 in the elution buffer. Storage Store at -80°C. Aliquot to avoid repeated freezing and thawing. Instruction: Note: Best use within three months from the date of receipt of this protein. MSDS: Download Datasheet: Download Applications Enzyme-linked Immunoabsorbent Assay Western Blot (Recombinant protein) Antibody Production Protein Array Page 1 of 2 2016/5/22 Gene Information Entrez GeneID: 8170 GeneBank NM_007163 Accession#: Protein NP_009094 Accession#: Gene Name: SLC14A2 Gene Alias: FLJ16167,HUT2,MGC119566,MGC119567,UT-A2,UT2,UTA,UTR,hUT- A6 Gene solute carrier family 14 (urea transporter), member 2 Description: Omim ID: 601611 Gene Ontology: Hyperlink Gene Summary: In mammalian cells, urea is the chief end-product of nitrogen catabolism and plays an important role in the urinary concentration
    [Show full text]