DOCSLIB.ORG

Comprehensive Profiling of Glycosphingolipid Glycans Using A

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Comprehensive Profiling of Glycosphingolipid Glycans Using A Article Comprehensive Profiling of Glycosphingolipid Glycans using a Novel Broad Specificity Endoglycoceramidase in a High-Throughput Workflow Simone Albrecht, Saulius Vainauskas, Henning Stoeckmann, Ciara McManus, Christopher H. Taron, and Pauline Mary Rudd Anal. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.analchem.6b00259 • Publication Date (Web): 01 Apr 2016 Downloaded from http://pubs.acs.org on April 6, 2016 Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts. Analytical Chemistry is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties. Page 1 of 22 Analytical Chemistry 1 2 3 4 Comprehensive Profiling of Glycosphingolipid Glycans using a Novel Broad Specificity 5 6 Endoglycoceramidase in a High-Throughput Workflow 7 8 9 Simone Albrecht 1, Saulius Vainauskas 2, Henning Stöckmann 1, 10 11 Ciara McManus 1, Christopher H. Taron 2Δ, Pauline M. Rudd 1Δ* 12 13 14 1NIBRT GlycoScience Group, National Institute for Bioprocessing, Research and Training, Fosters 15 16 Avenue, Mount Merrion, Blackrock, Dublin 4, Ireland 17 18 19 2New England Biolabs, Ipswich, MA, USA 20 21 22 23 24 ΔChristopher Taron and Pauline Rudd share senior authorship 25 26 27 28 *To whom correspondence should be addressed: Pauline M. Rudd, NIBRT GlycoScience Group, 29 National Institute for Bioprocessing, Research and Training, Fosters Avenue, Mount Merrion, 30 31 Blackrock, Dublin 4, Ireland. Tel.: +353 12158 142; Fax: +353 12158 116; E-mail: 32 33 [email protected] 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 1 ACS Paragon Plus Environment Analytical Chemistry Page 2 of 22 1 2 3 Abstract 4 5 The biological function of glycosphingolipids (GSLs) is largely determined by their glycan head 6 7 group moiety. This has placed a renewed emphasis on detailed GSL head group structural analysis. 8 Comprehensive profiling of GSL head groups in biological samples requires the use of 9 10 endoglycoceramidases with broad substrate specificity and a robust workflow that enables their 11 12 high-throughput analysis. We present here the first high-throughput glyco-analytical platform for 13 GSL head group profiling. The workflow features enzymatic release of GSL glycans with a novel 14 15 broad-specificity endoglycoceramidase I (EGCase I) from Rhodococcus triatomea , selective glycan 16 17 capture on hydrazide beads on a robotics platform, 2AB-fluorescent glycan labelling and analysis by 18 UPLC-HILIC-FLD. R. triatomea EGCase I displayed a wider specificity than known EGCases and was 19 20 able to efficiently hydrolyze gangliosides, globosides, (n)Lc-type GSLs and cerebrosides. Our 21 workflow was validated on purified GSL standard lipids and was applied to the characterization of 22 23 GSLs extracted from several mammalian cell lines and human serum. This study should facilitate 24 25 the analytical workflow in functional glycomics studies and biomarker discovery. 26 27 28 Keywords: , ultra-performance hydrophilic interaction liquid chromatography (UPLC-HILIC), 29 30 endoglycoceramidase, glycan profiling, glycosphingolipid, glycomics, high-throughput 31 32 33 34 35 Abbreviations: GSL, glycosphingolipid; MODY, maturity-onset diabetes of the young; rEGCase, 36 recombinant endoglycoceramidase; EGALC, endogalactosylceramidase; Gal, galactose; Cer, 37 38 ceramide; 2AB, 2-aminobenzamide; UPLC-HILIC-FLD, ultra performance liquid chromatography- 39 40 hydrophilic interaction-fluorescence detection; IPTG, isopropyl-β-thiogalactopyranoside; ACN, 41 acetonitrile; MeOH, methanol; GU, glucose units; QTOF, quadruploe time-of-flight; WAX, weak anion 42 43 exchange; ABS, A.ureafaciens α(2-3/6/8)-sialidase; NAN1, S. pneumoniae α(2-3)-sialidase; BKF, 44 45 bovine kidney α(1-2/4)-fucosidase; AMF, almond meal α(1-3/4)-fucosidase; BTG, bovine testes 46 β(1-3/4)-galactosidase; SPG, S. pneumoniae β(1-4)-galactosidase; CBG, coffee bean α(1-3/4)- 47 48 galactosidase; JBH, jack bean β(1-2/3/4/6) -N-acetylhexosaminidase; CV, coefficient of variance; 49 50 SSEA, stage specific embryonic antigen; HLB, hydrophilic-lipophilic balance; Neu5Ac/S, N- 51 acetylneuraminic acid; Neu5Gc/Sg, N-glycolylneuraminic acid; Fuc/F, fucose; Hex, hexose; HexNAc, 52 53 N-acetylhexosamine; GlcNAc, N-acetylglucosylamine. 54 55 56 57 58 59 60 2 ACS Paragon Plus Environment Page 3 of 22 Analytical Chemistry 1 2 3 Introduction 4 5 Many mammalian secretory proteins and lipids contain covalently linked carbohydrates (glycans). 6 7 For these molecules, the structure and composition of their appended glycans plays a significant 8 role in their function, distribution and physical properties 1. As glycan biosynthesis is not directly a 9 10 template-driven process, glycan structures on secreted glycoproteins and cell surface lipids are 11 12 typically heterogeneous and complex. Additionally, glycans are subject to structural alterations 13 over their lifetime due to both normal and pathological physiological processes, a feature that 14 15 makes them attractive as potential biomarkers of disease. 16 17 In the past few years, there have been major advances in high-throughput glycomics 18 technologies for glycoprotein analysis. A low-cost, high-throughput, automated N-glycan sample 19 20 preparation platform for glycoprofiling of immunoglobulins (IgG), antibodies and glycoproteins 21 isolated from serum was recently described by our laboratory 2-5. To date the analysis of N-glycans 22 23 has largely been based on chromatographic profiling using ultra-performance hydrophilic 24 25 interaction liquid chromatography with fluorescence detection (UPLC-HILIC-FLD). This technique 26 permits rapid and semi-quantitative comparison of N-glycan structures across many samples. 27 28 Serum N-glycan profiling has been used to identify glycan biomarkers of various diseases such as 29 30 cancer (ovarian, prostate, breast, lung, pancreatic, stomach), mature onset diabetes of the young 31 (MODY) , as well as biomarkers associated with normal physiological processes like aging 6-8. While 32 33 this young field has made significant advances, it is desirable to expand the breadth of these 34 35 profiling studies to other families of glycoproteins and other glycoconjugates, for example, 36 glycolipids. 37 38 One attractive class of molecule for glycoprofiling studies are glycosphingolipids (GSLs), 39 40 lipids that possess a carbohydrate head group consisting of mono- or oligosaccharides attached to 41 the lipids sphingosine or ceramide. More than 500 structural species that differ in their head group 42 43 glycan or fatty acid composition are known 9. GSL glycan head groups are associated with many 44 45 cellular processes such as cell differentiation, signalling and receptor functions for viruses, 46 antibodies or lectins 10 . GSLs are ubiquitous on cell membranes and also circulate in serum where 47 48 they are present in a free form or in complex with proteins 11 . Aberrant GSL glycosylation has been 49 50 repeatedly reported for different types of cancer including lung, breast, prostate and ovarian cancer 51 as well as brain tumors, multiple sclerosis, rheumatoid arthritis and lysosomal storage diseases 52 53 such as Gaucher’s and Fabry disease 12-15 . 54 55 In contrast to N-glycan profiling, high-throughput analysis of GSLs is still in its infancy. 56 Although methods have been described for the automated extraction of GSL from their matrix and 57 58 their subsequent characterization en masse using shotgun lipidomics 16 no such automated methods 59 60 3 ACS Paragon Plus Environment Analytical Chemistry Page 4 of 22 1 2 3 are currently available for the preparation and chromatographic analysis of GSL glycans. 4 5 Enzymatically released GSL head groups are commonly analyzed using MALDI-TOF MS 17,18 . 6 7 Enzymes often play a critical role in glycomics workflows such as the enzymatic release of GSL 8 glycans by endoglycoceramidases (EGCases) which is preferred over harsh chemical release 9 10 methods that can result in low glycan yields 19,20 . However,
Load more

Recommended publications
  • From Genes to Systems: New Global Strategies for the Characterization of NCL Biology ⁎ Anu Jalanko A, , Jaana Tyynelä B, Leena Peltonen A,C,D,E
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1762 (2006) 934–944 www.elsevier.com/locate/bbadis Review From genes to systems: New global strategies for the characterization of NCL biology ⁎ Anu Jalanko a, , Jaana Tyynelä b, Leena Peltonen a,c,d,e a National Public Health Institute, Department of Molecular Medicine, Biomedicum Helsinki, P.O. Box 104, 00251 Helsinki, Finland b University of Helsinki, Institute of Biomedicine/Biochemistry, Finland c University of Helsinki, Department of Medical Genetics, Finland d Research Programme of Molecular Medicine and Research Programme of Neurosciences, Biomedicum Helsinki, Finland e The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA Received 22 April 2006; received in revised form 1 September 2006; accepted 5 September 2006 Available online 12 September 2006 Abstract Neuronal ceroid lipofuscinoses (NCL) are rare neurological disorders with a uniform phenotype, caused by mutations in seven known genes. NCL provide a unique model to characterize molecular pathways critical for normal neuronal development and pathological neuronal degeneration. Systems biology based approach utilizes the rapidly developing tools of genomics, proteomics, lipidomics and metabolomics and aims at thorough understanding of the functions of cells, tissues and whole organisms by molecular analysis and biocomputing-assisted modeling. The systems level understanding of NCL is now possible by utilizing different model organisms. Initial work has revealed disturbed metabolic pathways in several NCL disorders and most analyses have utilized the infantile (INCL/CLN1) and juvenile (JNCL/CLN3) disease modeling and utilized mainly human and mouse samples.
    [Show full text]
  • Sphingolipid Metabolism Diseases ⁎ Thomas Kolter, Konrad Sandhoff
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1758 (2006) 2057–2079 www.elsevier.com/locate/bbamem Review Sphingolipid metabolism diseases ⁎ Thomas Kolter, Konrad Sandhoff Kekulé-Institut für Organische Chemie und Biochemie der Universität, Gerhard-Domagk-Str. 1, D-53121 Bonn, Germany Received 23 December 2005; received in revised form 26 April 2006; accepted 23 May 2006 Available online 14 June 2006 Abstract Human diseases caused by alterations in the metabolism of sphingolipids or glycosphingolipids are mainly disorders of the degradation of these compounds. The sphingolipidoses are a group of monogenic inherited diseases caused by defects in the system of lysosomal sphingolipid degradation, with subsequent accumulation of non-degradable storage material in one or more organs. Most sphingolipidoses are associated with high mortality. Both, the ratio of substrate influx into the lysosomes and the reduced degradative capacity can be addressed by therapeutic approaches. In addition to symptomatic treatments, the current strategies for restoration of the reduced substrate degradation within the lysosome are enzyme replacement therapy (ERT), cell-mediated therapy (CMT) including bone marrow transplantation (BMT) and cell-mediated “cross correction”, gene therapy, and enzyme-enhancement therapy with chemical chaperones. The reduction of substrate influx into the lysosomes can be achieved by substrate reduction therapy. Patients suffering from the attenuated form (type 1) of Gaucher disease and from Fabry disease have been successfully treated with ERT. © 2006 Elsevier B.V. All rights reserved. Keywords: Ceramide; Lysosomal storage disease; Saposin; Sphingolipidose Contents 1. Sphingolipid structure, function and biosynthesis ..........................................2058 1.1.
    [Show full text]
  • Glycosphingolipid Metabolism in Cell Fate Specification Domenico Russo1,*, Laura Capolupo1,2, Jaipreet Singh Loomba1,2, Lucia Sticco1 and Giovanni D’Angelo1,2,*
    © 2018. Published by The Company of Biologists Ltd | Journal of Cell Science (2018) 131, jcs219204. doi:10.1242/jcs.219204 REVIEW Glycosphingolipid metabolism in cell fate specification Domenico Russo1,*, Laura Capolupo1,2, Jaipreet Singh Loomba1,2, Lucia Sticco1 and Giovanni D’Angelo1,2,* ABSTRACT anchoring. Specifically at the PM, sphingolipids participate in Glycosphingolipids (GSLs) are ubiquitous components of eukaryotic signaling events by recruiting signaling molecules to, or plasma membranes that consist of a ceramide backbone linked to a sequestering them at, membrane microdomains for the modulation glycan moiety. Both the ceramide and the glycan parts of GSLs display of their activities and for their processing into the endocytic cycle structural variations that result in a remarkable repertoire of diverse (Holthuis and Menon, 2014; Holthuis et al., 2001; Simons and compounds. This diversity of GSLs is exploited during embryogenesis, Ikonen, 1997). Given these properties, sphingolipids are proposed to when different GSLs are produced at specific developmental stages and function as fundamental membrane organizers and to make up along several differentiation trajectories. Importantly, plasma membrane the fabric of eukaryotic PMs in order to influence the interaction receptors interact with GSLsto modify their activities. Consequently, two with the extracellular environment (Hannun and Obeid, 2018; otherwise identical cells can respond differently to the same stimulus Holthuis et al., 2001). owing to their different GSL composition. The metabolic reprograming of Interestingly, different cell types exhibit a specific sphingolipid GSLs is in fact a necessary part of developmental programs, as its array at their PMs (Hakomori, 2003; Ngamukote et al., 2007) impairment results in developmental failure or tissue-specific defects.
    [Show full text]
  • Building a Metabolic Bridge Between Glycolysis and Sphingolipid Biosynthesis : Implications in Cancer
    University of Louisville ThinkIR: The University of Louisville's Institutional Repository Electronic Theses and Dissertations 8-2014 Building a metabolic bridge between glycolysis and sphingolipid biosynthesis : implications in cancer. Morgan L. Stathem University of Louisville Follow this and additional works at: https://ir.library.louisville.edu/etd Part of the Pharmacy and Pharmaceutical Sciences Commons Recommended Citation Stathem, Morgan L., "Building a metabolic bridge between glycolysis and sphingolipid biosynthesis : implications in cancer." (2014). Electronic Theses and Dissertations. Paper 1374. https://doi.org/10.18297/etd/1374 This Master's Thesis is brought to you for free and open access by ThinkIR: The nivU ersity of Louisville's Institutional Repository. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of ThinkIR: The nivU ersity of Louisville's Institutional Repository. This title appears here courtesy of the author, who has retained all other copyrights. For more information, please contact [email protected]. BUILDING A METABOLIC BRIDGE BETWEEN GLYCOLYSIS AND SPHINGOLIPID BIOSYNTHESIS: IMPLICATIONS IN CANCER By Morgan L. Stathem B.S., University of Georgia, 2010 A Thesis Submitted to the Faculty of the School of Medicine of the University of Louisville In Partial Fulfillment of the Requirements for the Degree of Master of Science Department of Pharmacology and Toxicology University of Louisville Louisville, KY August 2014 BUILDING A METABOLIC BRIDGE BETWEEN GLYCOLYSIS AND SPHINGOLIPID BIOSYNTHESIS: IMPLICATIONS IN CANCER By Morgan L. Stathem B.S., University of Georgia, 2010 Thesis Approved on 08/07/2014 by the following Thesis Committee: __________________________________ Leah Siskind, Ph.D. __________________________________ Levi Beverly, Ph.D.
    [Show full text]
  • Download (Accessed on 10 December 2020)
    H OH metabolites OH Article Comprehensive Identification of Glycosphingolipids in Human Plasma Using Hydrophilic Interaction Liquid Chromatography— Electrospray Ionization Mass Spectrometry Karel Hoˇrejší 1,2, Robert Jirásko 1, Michaela Chocholoušková 1, Denise Wolrab 1, David Kahoun 2 and Michal Holˇcapek 1,* 1 Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 532 10 Pardubice, Czech Republic; [email protected] (K.H.); [email protected] (R.J.); [email protected] (M.C.); [email protected] (D.W.) 2 Institute of Chemistry, Faculty of Science, University of South Bohemia in Ceskˇ é Budˇejovice,Branišovská 1760, 370 05 Ceskˇ é Budˇejovice,Czech Republic; [email protected] * Correspondence: [email protected]; Tel.: +420-466-037-087 Abstract: Glycosphingolipids (GSL) represent a highly heterogeneous class of lipids with many cellular functions, implicated in a wide spectrum of human diseases. Their isolation, detection, and comprehensive structural analysis is a challenging task due to the structural diversity of GSL molecules. In this work, GSL subclasses are isolated from human plasma using an optimized monophasic ethanol–water solvent system capable to recover a broad range of GSL species. Obtained Citation: Hoˇrejší,K.; Jirásko, R.; deproteinized plasma is subsequently purified and concentrated by C18-based solid-phase extraction Chocholoušková, M.; Wolrab, D.; (SPE). The hydrophilic interaction liquid chromatography coupled to electrospray ionization linear Kahoun, D.; Holˇcapek,M. ion trap tandem mass spectrometry (HILIC-ESI-LIT-MS/MS) is used for GSL analysis in the human Comprehensive Identification of plasma extract. Our results provide an in-depth profiling and structural characterization of glycosph- Glycosphingolipids in Human ingolipid and some phospholipid subclasses identified in the human plasma based on their retention Plasma Using Hydrophilic Interaction times and the interpretation of tandem mass spectra.
    [Show full text]
  • Anti-GM2 Ganglioside [L55-81] Standard Size, 200 Μg, Ab00786-23.0 View Online
    Anti-GM2 ganglioside [L55-81] Standard Size, 200 μg, Ab00786-23.0 View online Anti-GM2 ganglioside [L55-81] Standard Size Ab00786-23.0 This chimeric rabbit antibody was made using the variable domain sequences of the original Human IgM format, for improved compatibility with existing reagents, assays and techniques. Isotype and Format: Rabbit IgG, Kappa Clone Number: L55-81 Alternative Name(s) of Target: UniProt Accession Number of Target Protein: Published Application(s): depleting, TLC, ELISA, IHC Published Species Reactivity: Human Immunogen: L55-81 was prepared by isolating EBV-immortalized peripheral blood B lymphocytes from a human donor with melanoma. Specificity: L55-81 binds specifically to GM2 ganglioside; this antibody shows no reactivity towards GM1, GM3, GD2 or normal tissues- the L55-81 epitope contains both a GalNAc and sialic acid bound to lactose. Ganglioside GM2 (GalNAcβ1-4(NeuNAcα2-3)Galβ1-4Glcβ-1ceramide) is a glycosphingolipid present on the cell surface and a major tumor-differentiation antigen in human melanoma and hepatoma. Application Notes: L55-81 reacts positively to melanoma cell lines with high GM2 expression (M14; M24), and shows no binding to normal cell types or cell lines M15 or M25 which have low GM2 expression and high GM3 expression. This specificity can be confirmed by ELISA and TLC immunostaining by testing a variety of gangliosides and neutral glycolipids. L55-81 is able to lyse cells from the M14 melanoma cell line (which has high GM2 expression) in the presence of human or rabbit complement, but not in the absence of complement - this was shown by a [51Cr] release assay.
    [Show full text]
  • The Glycosphingolipid, Lactosylceramide, Regulates B1-Integrin Clustering and Endocytosis
    Research Article The Glycosphingolipid, Lactosylceramide, Regulates B1-Integrin Clustering and Endocytosis Deepak K. Sharma, Jennifer C. Brown, Zhijie Cheng, Eileen L. Holicky, David L. Marks, and Richard E. Pagano Department of Biochemistry and Molecular Biology, Thoracic Diseases Research Unit, and Mayo Clinic Cancer Center, Mayo Clinic College of Medicine, Rochester, Minnesota Abstract signaling complexes (3, 5) and are also initiation sites for clathrin- independent endocytic events (3, 5, 6). Glycosphingolipids are known to play roles in integrin- mediated cell adhesion and migration; however, the mecha- One mechanism by which glycosphingolipids could affect cell nisms by which glycosphingolipids affect integrins are adhesion and migration is via their interaction with integrins. ah unknown. Here, we show that addition of the glycosphingolipid, Integrins are a family of heterodimeric, integral membrane C8-lactosylceramide (C8-LacCer), or free cholesterol to human proteins at the plasma membrane, which bind to extracellular fibroblasts at 10°C causes the formation of glycosphingolipid- matrix (ECM) proteins and cell surface ligands, and are responsible enriched plasma membrane domains as shown by visualizing a for many types of cell adhesion events (7, 8). Glycosphingolipids have fluorescent glycosphingolipid probe, BODIPY-LacCer, incorpo- been shown to directly modulate integrin-based cell attachment. For rated into the plasma membrane of living cells. Addition of C8- example, gangliosides (sialic acid–terminated glycosphingolipids) LacCer or cholesterol to cells initiated the clustering of extracted from neuroblastoma cells or atherosclerotic plaques enhance platelet adhesion via integrin binding to collagen (9–11). B -integrins within these glycosphingolipid-enriched domains 1 Gangliosides also enhance binding of integrins to the ECM in mouse and the activation of the B1-integrins as assessed using a HUTS antibody that only binds activated integrin.
    [Show full text]
  • Tay-Sachs Disease Enfermedad De Tay-Sachs Received: 10/01/2018
    Rev. Fac. Med. 2019 Vol. 67 No. 3: 511-7 511 REVIEW ARTICLE DOI: http://dx.doi.org/10.15446/revfacmed.v67n3.69742 Tay-Sachs disease Enfermedad de Tay-Sachs Received: 10/01/2018. Accepted: 14/06/2018. Carlos Andrés Gualdrón-Frías1 • Laura Tatiana Calderón-Nossa1 1 Universidad Pedagógica y Tecnológica de Colombia - Tunja Campus - Faculty of Health Sciences - School of Medicine - ACEMED-UPTC Research Group - Tunja - Colombia Corresponding author: Carlos Andrés Gualdrón-Frías. Grupo de Investigación ACEMED-UPTC, Facultad de Ciencias de la Salud, Universidad Pedagógica y Tecnológica de Colombia. Calle 24 No. 5-63, antiguo Hospital San Rafael de Tunja. Phone number: +57 8 7405626, ext.: 2908. Tunja. Colombia. Email: [email protected]. | Abstract | | Resumen | Introduction: Lysosomal storage disease is caused by the deficiency Introducción. La deficiencia de una sola hidrolasa (enzimas lisosomales) of a single hydrolase (lysosomal enzymes). GM2 gangliosidoses da como resultado una enfermedad de almacenamiento lisosomal. Las are autosomal recessive disorders caused by deficiency of gangliosidosis GM2 son trastornos autosómicos recesivos causados β-hexosaminidase and Tay-Sachs disease (TSD) is one of its three por la deficiencia de β-hexosaminidasa. La enfermedad de Tay-Sachs forms. (TSD, por sus siglas en inglés) es una de las tres presentaciones de este tipo de gangliosidosis. Objective: To perform a review of the state of the art on TSD and describe its definition, epidemiology, etiology, physiopathology, Objetivo. Realizar una revisión del estado del arte de la TSD describiendo clinical manifestations, as well as advances regarding its diagnosis su definición, epidemiología, etiología, fisiopatología, manifestaciones and treatment. clínicas y actualidades en su diagnóstico y tratamiento.
    [Show full text]
  • A Role for Glycosphingolipid Accumulation in the Renal Hypertrophy of Streptozotocin-Induced Diabetes Mellitus
    A role for glycosphingolipid accumulation in the renal hypertrophy of streptozotocin-induced diabetes mellitus. I Z Zador, … , N S Radin, J A Shayman J Clin Invest. 1993;91(3):797-803. https://doi.org/10.1172/JCI116299. Research Article Glucosylceramide (GlcCer) and related glycosphingolipids have been implicated as causal elements in both the growth of cells and in the regulation of hormonal signaling. We therefore studied whether the renal hypertrophy induced by diabetes was associated with enhanced synthesis of glycosphingolipids. 16 d after the induction of diabetes, increases in renal size and concentration of glucocerebroside and ganglioside GM3 were observed paralleling an increase in UDP-Glc concentration. GlcCer synthase and beta-glucosidase-specific activities were no different between control and diabetic kidneys. The apparent Km of the GlcCer synthase with respect to UDP-Glc was 250 microM and was unchanged in the diabetic kidneys. The observed concentrations of UDP-Glc were 149 and 237 microM in control and diabetic kidneys, respectively. The UDP-Glc level is thus rate limiting with regard to GlcCer synthesis. To determine whether the changes in glycolipid content were functionally significant, diabetic and control groups were treated with the GlcCer synthase inhibitor, D-threo-1-phenyl-2-decanoyl-amino-3-morpholino-1- propanol, 2 wk after the induction of diabetes. Kidney weights in the diabetic rats treated with D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol were no different than the control groups. Morphometric analysis of glomerular volumes paralleled changes in renal growth. Glycosphingolipid formation may therefore represent a significant pathway for glucose utilization in early diabetic nephropathy.
    [Show full text]
  • Investigating Sphingolipid Behavior and Function Using Metabolic Labeling
    INVESTIGATING SPHINGOLIPID BEHAVIOR AND FUNCTION USING METABOLIC LABELING BY RAEHYUN KIM DISSERTATION Submitted in partial fulfillment of the requirements for degree of Doctor of Philosophy in Chemical Engineering in the Graduate College of the University of Illinois at Urbana-Champaign, 2016 Urbana, Illinois Doctoral Committee: Associate Professor Mary L. Kraft, Chair Professor Ryan C. Bailey Professor Deborah E. Leckband Associate Professor Charles M. Schroeder ABSTRACT The past few decades of research have accumulated a body of evidence that membrane lipids are far more than merely the structural components of biological membranes. Instead, membrane lipids play important roles in cellular functions in multiple ways. Sphingolipids are a group of lipids that are involved in various cellular processes that are crucial for cell survival and proliferation. However, our understanding of sphingolipid function is limited due to the complexity of their behaviors and the lack of proper tools to address and decipher this complexity. Chapters 2 and 3 present metabolic labeling with fluorophores and stable isotope tags, respectively, as tools to investigate sphingolipid behaviors. Metabolic labeling enables one to detect and directly observe sphingolipids, and not the activities or levels of the enzymes that metabolize them. Metabolic labeling of cells with fluorescent sphingosines enabled visualization of the sphingosine metabolites in live cells and also showed potential for studies of metabolism and in vitro assays. Use of stable isotope tagged sphingolipid precursors, in conjunction with LC-MS/MS analysis, provided a more comprehensive and complete dataset than traditional radiolabeling, including information about unlabeled as well as labeled species. These tools offer great opportunities to explore sphingolipid behaviors.
    [Show full text]
  • Targeting Stable Nucleic Acid Lipid Particle (SNALP)-Formulated Sirnas to the Brain As a Therapeutic Approach
    International Journal of Molecular Sciences Review Lysosomal Storage Disease-Associated Neuropathy: Targeting Stable Nucleic Acid Lipid Particle (SNALP)-Formulated siRNAs to the Brain as a Therapeutic Approach Maria Francisca Coutinho 1,2,* , Juliana Inês Santos 1,3, Liliana S. Mendonça 4,5, Liliana Matos 1,2, Maria João Prata 3,6 , Amália S. Jurado 7, Maria C. Pedroso de Lima 4 and Sandra Alves 1,2 1 Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge (INSA I.P), Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal; [email protected] (J.I.S.); [email protected] (L.M.); [email protected] (S.A.) 2 Center for the Study of Animal Science, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal 3 Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal; [email protected] 4 CNC—Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal; [email protected] (L.S.M.); [email protected] (M.C.P.d.L.) 5 CIBB—Center for Innovative Biomedicine and Biotechnology, University of Coimbra, 3004-504 Coimbra, Portugal 6 i3S—Institute of Research and Innovation in Health/IPATIMUP—Institute of Molecular Pathology and Immunology of the University of Porto, Rua Alfredo Allen, 208 4200-135 Porto, Portugal 7 University of Coimbra, CNC—Center for Neuroscience and Cell Biology, Department of Life Sciences, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal; [email protected] * Correspondence: [email protected]; Tel.: +351-(223)-401-113 Received: 6 July 2020; Accepted: 6 August 2020; Published: 10 August 2020 Abstract: More than two thirds of Lysosomal Storage Diseases (LSDs) present central nervous system involvement.
    [Show full text]
  • Function of Platelet Glycosphingolipid Microdomains/Lipid Rafts
    International Journal of Molecular Sciences Review Function of Platelet Glycosphingolipid Microdomains/Lipid Rafts Keisuke Komatsuya , Kei Kaneko and Kohji Kasahara * Laboratory of Biomembrane, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan; [email protected] (K.K.); [email protected] (K.K.) * Correspondence: [email protected] Received: 29 June 2020; Accepted: 30 July 2020; Published: 2 August 2020 Abstract: Lipid rafts are dynamic assemblies of glycosphingolipids, sphingomyelin, cholesterol, and specific proteins which are stabilized into platforms involved in the regulation of vital cellular processes. The rafts at the cell surface play important functions in signal transduction. Recent reports have demonstrated that lipid rafts are spatially and compositionally heterogeneous in the single-cell membrane. In this review, we summarize our recent data on living platelets using two specific probes of raft components: lysenin as a probe of sphingomyelin-rich rafts and BCθ as a probe of cholesterol-rich rafts. Sphingomyelin-rich rafts that are spatially and functionally distinct from the cholesterol-rich rafts were found at spreading platelets. Fibrin is translocated to sphingomyelin-rich rafts and platelet sphingomyelin-rich rafts act as platforms where extracellular fibrin and intracellular actomyosin join to promote clot retraction. On the other hand, the collagen receptor glycoprotein VI is known to be translocated to cholesterol-rich rafts during platelet adhesion to collagen. Furthermore, the functional roles of platelet glycosphingolipids and platelet raft-binding proteins including G protein-coupled receptors, stomatin, prohibitin, flotillin, and HflK/C-domain protein family, tetraspanin family, and calcium channels are discussed. Keywords: lipid rafts; detergent-resistant membrane; heterogeneity; platelets; lysenin 1.
    [Show full text]