Glycoproteins and N-glycans from exosomes
|
|
- Sydney Sherman
- 5 years ago
- Views:
Transcription
1 Glycoproteins and N-glycans from exosomes Júlia Costa Laboratory of Glycobiology WP3: Exosome specific glycosignatures defining specificity in exosomes targeting GlioEx University Medical Center Hamburg Eppendorf 22 May 2014
2 The erythrocyte glycocalyx Roseman, 2001
3 Localizations and functions of glycoconjugates Blood and other fluids - Solubility - Stability - Recognition by carbohydrate-binding proteins Golgi apparatus Bacterial infection Cell-cell interactions - Metastasis - Inflammation Lysosomal targeting (Man6P) Nucleus Endoplasmic reticulum Lysosome Viral infection Quality control of glycoprotein folding Luminal/extracellular glycans Cytoplasmic/nuclear glycans Cell-matrix interactions Carrier proteins or lipids Glycan-binding proteins Extracellular matrix
4 N-glycosylation In N-glycosylation, glycans are covalently bound to asparagine side chains (NXS/T, X P) of proteins. In animal cells, the sugar linked to the asparagine residue is N-acetylglucosamine. trans-golgi and trans-golgi network Endoplasmic reticulum cis-golgi medial-golgi Oligosaccharyltransferase ER glucosidases I, II ER mannosidase Golgi mannosidases IA, IB, IC GlcNActransferase I Golgi mannosidase II GlcNActransferase II Fucosyltransferase VIII Galactosyltransferase Sialyltransferases Peripheral fucosyltransferases Mannose Glucose N-Acetylglucosamine Galactose Fucose N-Acetylneuraminic acid After Taylor and Drickamer, 2006
5 O-glycosylation In O-glycosylation glycans are covalently bound to the side chains of serine, threonine, tyrosine, hydroxylysine, or hydroxyproline side-chains. - Mucins - Proteoglycans - O-linked Man - O-linked GlcNAc
6 Examples of peripheral glycosylation linked to the LacNAc structure α2,6 α1,2 α1,3/4 6 SO 3 - α2,8 α2,3 β1,4 β1-r β1-4 α1,3 β1,4 N-Acetylglucosamine Galactose Fucose N-Acetylneuraminic acid
7 Multiple approaches for profiling a cell s glycome Derivatization Liquid chromatography Mass spectrometry (2009) Essentials of Glycobiology
8 Exosomes Théry et al. (2009) Nat. Rev. Immunol. 9, 581
9 Exosome separation Ovarian carcinoma SKOV3 cells 150 µm Centrifugation of cell supernatant at 500xg, 10,000xg, and 100,000xg (crude exosomes) 100,000xg pellet Escrevente et al. (2008) BBA 1780, 905
10 Exosome separation Exosc Exos 100 nm 100 nm Escrevente et al. (2013) PLOS One 8, e78631
11 Structural analysis of protein glycosylation Lectin blotting Immunoblotting Exosomes Glycoproteins Lectin affinity chromatography Peptide mass fingerprinting Glycan structural analysis Glycan release from tryptic glycopeptides: peptide N-glycosidase F (PNGase F) Glycan labelling: 2-aminobenzamide (2-AB) Glycan separation and structure elucidation: HPAEC-PAD, NP-HPLC, exoglycosidase digestion, mass spectrometry
12 Sialoglycoproteins from exosomes Ovarian carcinoma cell lines SKOV3 kda Ext Exos c Ext Exos c Ext Exos c Ext Exos c Ext Exos c Ext Exos c Ext Exos c Ext Exos c Embryonic kidney HEK293 Neuroglioma H4 MAL - NeuAcα2,3Gal; SNA - NeuAcα2,6Gal/GalNAc; kda Ext Exos c Ext Exos c kda Ext Exos c Ext Exos c Escrevente et al. (2011) BMC Can. 11, 108
13 Exosomes have specific glycosylation profiles in comparison with plasma membrane and microsomal fractions Ovarian carcinoma SKOV3 cells kda Ext MF PM IC Cyt Exos Ves c Maackia amurensis lectin (MAL) specificity: NeuAcα2,3Gal Sambucus nigra aglutinin (SNA) specificity: NeuAcα2,6Gal/GalNAc Wisteria floribunda (WFA) specificity: GalNAcβ4GlcNAc (LacdiNAc) Concanavalin A (Con A) specificity: α-mannosyl containing branched glycans
14 Sialoglycoproteins from exosomes Ovarian carcinoma SKOV3 cells Exosomes kda WB: CD9
15 MAL-binding glycoproteins from exosomes MAL-affinity purified IP: anti-lgals3bp kda Exos c -Ab -Exos c IP MAL Galectin-3 binding protein LGALS3BP Bands 4-8, 10 identified by a proteomics approach (peptide mass fingerprinting, LC-MS/MS) as galectin-3 binding protein. LGALS3BP is a secreted glycoprotein that has been found in the extracellular matrix and in body fluids and exosomes It has been found in several cancers where it has a negative prognostic value and is involved in cell adhesion
16 NP-HPLC of 2-AB-labeled N-glycans after PNGase F release 7x10 5 M5 M6 M7 M8 M9 Exosomes High mannose glycans 0 5x x M5 M5 M6 M7 M8 M9 M6 M7 M8 M9 Microsomal fraction Plasma membrane Mannose N-Acetylglucosamine Minutes Exosomes (23%) Microsomal fraction (41%) Plasma membrane (15%) Exosomes, microsomal fraction and plasma membrane have distinct N-glycosylation signatures.
17 Digestion of N-glycans from exosomes with exoglycosidases M: α-(1-2,3,6)-mannosidase S: α-(2-3/6)-sialidase G: β(1-3,4)-galactosidase F: α-l-(1-2,3,4,6)-fucosidase Sialic acid containing complex-type N-glycans Exosomes (77%) (approx NeuAc content: 170 pmol/µg total protein) Microsomal fraction (35%) Plasma membrane (26%)
18 Glycosidases used for structure analysis
19 MALDI-TOF MS of N-glycans Intens. 600 * * Exosomes Microsomal fraction Plasma membrane m/z *LC-ESI Ion Trap MS analyses; collision induced fragmentation; fragments identified corresponding to bisecting GlcNAc
20 Conclusions and perspectives Exosomes have characteristic glycan signatures. The sialoglycoprotein LGALS3BP is an abundant exosome marker. Does LGALS3BP and specific glycans play a role in interaction with target cells? Engineering of exosomes with respect to glycoproteins/glycosylation may improve their interaction with target cells. SKOV3 cell after internalization of CSFE labeled exosomes Escrevente (2012) BMC Can 11, 108
21 Acknowledgements Laboratory of Glycobiology Present: Margarida Gonçalves Gonçalo Silva Teresa Silva Former post-docs and students: Stefanie Glathe, Francisco Enguita, Carla Jorge, Vitor Sousa, Vanessa Morais, Angelina Palma, Catarina Brito, Catarina Gomes, Eda Machado, Rita Carilho, Ricardo Gouveia, Cristina Escrevente, Adriana Gomes, Joana Batista, Susana Jorge Collaborations in exosomes projects Harald Conradt, GlycoThera, Hannover Peter Altevogt, DKFZ, Heidelberg Paula Alves, Patrícia Alves, ITQB, IBET, Oeiras Funding (past 5 years) FCT, Portugal: PTDC/SAU-NEU/100724/2008; PIC/IC/82765/2007; SOPHIA, JPND/0003/2011 (EU Joint Programme Neurodegenerative Disease Research); GlioEx ENMed/0001/2013 (EuroNanomedII); PhD fellowships
Nature Biotechnology: doi: /nbt Supplementary Figure 1. RNAseq expression profiling of selected glycosyltransferase genes in CHO.
Supplementary Figure 1 RNAseq expression profiling of selected glycosyltransferase genes in CHO. RNAseq analysis was performed on two common CHO lines (CHO-K1, CHO-GS) and two independent CHO-GS triple
More informationSignificance and Functions of Carbohydrates. Bacterial Cell Walls
Biochemistry 462a - Carbohydrate Function Reading - Chapter 9 Practice problems - Chapter 9: 2, 4a, 4b, 6, 9, 10, 13, 14, 15, 16a, 17; Carbohydrate extra problems Significance and Functions of Carbohydrates
More informationBiosynthesis of N and O Glycans
TechNote #TNGL101 Biosynthesis of N and O Glycans These suggestions and data are based on information we believe to be reliable. They are offered in good faith, but without guarantee, as conditions and
More informationEFFECT OF VITAMINS C AND E ON THE LEVELS OF CARBOHYDRATE COMPONENTS OF GLYCOPROTEINS IN COLLAGEN INDUCED ARTHRITIS
Chapter 4 EFFECT OF VITAMINS C AND E ON THE LEVELS OF CARBOHYDRATE COMPONENTS OF GLYCOPROTEINS IN COLLAGEN INDUCED ARTHRITIS 4.1. INTRODUCTION Rheumatoid Arthritis is a chronic inflammatory disease categorized
More informationEnvelope glycans of immunodeficiency virions are almost entirely oligomannose antigens
Supporting Information for: Envelope glycans of immunodeficiency virions are almost entirely oligomannose antigens Katie J. Doores *1,2, Camille Bonomelli *3, David J. Harvey 3, Snezana Vasiljevic 3, Raymond
More informationBIOSYNTHESIS OF CANCER-RELATED CARBOHYDRATE ANTIGENS. Fabio Dall Olio Department of Experimental Pathology University of Bologna, Italy
BIOSYNTHESIS OF CANCER-RELATED CARBOHYDRATE ANTIGENS Fabio Dall Olio Department of Experimental Pathology University of Bologna, Italy TOPICS OF THE LECTURE 1. Structure and function of some representative
More informationDr Mark Hilliard, NIBRT. Waters THE SCIENCE OF WHAT S POSSIBLE TM
RFMS Glycan Characterization Techniques for Biotherapeutics Dr Mark Hilliard, NIBRT Waters THE SCIENCE OF WHAT S POSSIBLE TM The Complexity of Glycosylation Glycosylation is the most common posttranslational
More informationGlycan and Monosaccharide Workshop Eoin Cosgrave David Wayland Bill Warren
Glycan and Monosaccharide Workshop Eoin Cosgrave David Wayland Bill Warren 2012 Waters Corporation 1 Requests and Questions Optimised sample prep protocol to reduce sample preparation time How can I detect
More informationUNIVERSITY OF YORK BIOLOGY. Glycobiology
Examination Candidate Number: This paper has two parts: UNIVERSITY OF YORK BSc Stage 3 Degree Examinations 2017-18 Department: BIOLOGY Title of Exam: Glycobiology Time allowed: 2 hours Total marks available
More informationWhat sort of Science is Glycoscience? (Introductory lecture)
Glycosciences: Glycobiology & Glycochemistry e-learning course What sort of Science is Glycoscience? (Introductory lecture) Paula Videira Faculdade de Ciências Médicas Nova University, Lisbon Portugal
More informationEnzymatic Removal of N- and O-glycans using PNGase F or the Protein Deglycosylation Mix
be INSPIRED drive DISCOVERY stay GENUINE APPLICATION NOTE Enzymatic Removal of N- and O-glycans using PNGase F or the Protein Deglycosylation Mix Alicia Bielik and Paula Magnelli, New England Biolabs,
More informationGlycosylation analyses of recombinant proteins by LC-ESI mass spectrometry
Glycosylation analyses of recombinant proteins by LC-ESI mass spectrometry Dr Malin Bäckström Mammalian Protein Expression Core Facility P4EU meeting Porto Nov 11-12, 2013 MPE - A tissue culture facility
More informationGlycoproteins and Mucins. B.Sopko
Glycoproteins and Mucins B.Sopko Content Glycoproteins: Structures and Linkages Interconversions and activation of dietary sugars Other pathways of sugar nucleotide metabolism Biosynthesis of oligosaccharides
More informationStructural Elucidation of N-glycans Originating From Ovarian Cancer Cells Using High-Vacuum MALDI Mass Spectrometry
PO-CON1347E Structural Elucidation of N-glycans Originating From Ovarian Cancer Cells Using High-Vacuum MALDI Mass Spectrometry ASMS 2013 TP-708 Matthew S. F. Choo 1,3 ; Roberto Castangia 2 ; Matthew E.
More informationTECHNICAL BULLETIN. Enzymatic Protein Deglycosylation Kit. Catalog Number EDEGLY Storage Temperature 2 8 C
Enzymatic Protein Deglycosylation Kit Catalog Number EDEGLY Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description The EDEGLY kit contains all the enzymes and reagents needed to completely remove
More informationTECHNICAL BULLETIN. R 2 GlcNAcβ1 4GlcNAcβ1 Asn
GlycoProfile II Enzymatic In-Solution N-Deglycosylation Kit Product Code PP0201 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description Glycosylation is one of the most common posttranslational
More informationCertificate of Analysis
Certificate of Analysis Human IgG Glycoprotein Standard Cat. #: GCP-IGG-50U Batch: B13T-06 Nominal size: 50μg Expiry: Dec 2020 Description: A glycoprotein standard for use during glycan release and labeling.
More informationChapter 11. Learning objectives: Structure and function of monosaccharides, polysaccharide, glycoproteins lectins.
Chapter 11 Learning objectives: Structure and function of monosaccharides, polysaccharide, glycoproteins lectins. Carbohydrates Fuels Structural components Coating of cells Part of extracellular matrix
More informationSupporting Information for MassyTools-assisted data analysis of total serum N-glycome changes associated with pregnancy
Supporting Information for MassyTools-assisted data analysis of total serum N-glycome changes associated with pregnancy Bas C. Jansen 1, Albert Bondt 1,2, Karli R. Reiding 1, Coen J. de Jong 1, David Falck
More informationChapter 10apter 9. Chapter 10. Summary
Chapter 10apter 9 Chapter 10 The field of proteomics has developed rapidly in recent years. The essence of proteomics is to characterize the behavior of a group of proteins, the system rather than the
More informationThank you for joining us! Our session will begin shortly Waters Corporation 1
UPLC and HPLC Separation Strategies for Successful Characterization of Glycans Derived from Therapeutic Proteins Thank you for joining us! Our session will begin shortly 2013 Waters Corporation 1 Friendly
More informationProtein Trafficking in the Secretory and Endocytic Pathways
Protein Trafficking in the Secretory and Endocytic Pathways The compartmentalization of eukaryotic cells has considerable functional advantages for the cell, but requires elaborate mechanisms to ensure
More informationBlotGlyco Glycan purification and labeling kit
Aug. 14th, 2012 BlotGlyco Glycan purification and labeling kit S-BIO Business Division SUMITOMO BAKELITE CO., LTD. 1 BlotGlyco kit Biological sample Glycan Hz BlotGlyco :Label MS HPLC 2 BlotGlyco kit is
More informationSummary of Endomembrane-system
Summary of Endomembrane-system 1. Endomembrane System: The structural and functional relationship organelles including ER,Golgi complex, lysosome, endosomes, secretory vesicles. 2. Membrane-bound structures
More informationSupplementary Figure 1. PD-L1 is glycosylated in cancer cells. (a) Western blot analysis of PD-L1 in breast cancer cells. (b) Western blot analysis
Supplementary Figure 1. PD-L1 is glycosylated in cancer cells. (a) Western blot analysis of PD-L1 in breast cancer cells. (b) Western blot analysis of PD-L1 in ovarian cancer cells. (c) Western blot analysis
More informationOligosaccharide Analysis by High-Performance Anion- Exchange Chromatography with Pulsed Amperometric Detection
Oligosaccharide Analysis by High-Performance Anion- Exchange Chromatography with Pulsed Amperometric Detection Jeff Rohrer, Ph.D. Director, Applications Development, Dionex Products 1 The world leader
More informationA. Incorrect! No, this is not the description of this type of molecule. B. Incorrect! No, this is not the description of this type of molecule.
Biochemistry - Problem Drill 08: Carbohydrates No. 1 of 10 1. have one aldehyde (-CHO) or one keto (-C=O) group and many hydroxyl (-OH) groups. (A) Amino acids (B) Proteins (C) Nucleic Acids (D) Carbohydrates
More informationThe use of mass spectrometry for the proteomic analysis of glycosylation
Proteomics 2006, 6, 3993 4015 DOI 10.1002/pmic.200600129 3993 REVIEW The use of mass spectrometry for the proteomic analysis of glycosylation Willy Morelle, Kévin Canis, Frédéric Chirat, Valegh Faid and
More informationN-Glycan Sequencing Kit
PROTEIN TOOLS N-Glycan Sequencing Kit Instruction Manual NEB #E577S 2 reactions Version 1. 1/18 be INSPIRED drive DISCOVERY stay GENUINE This product is intended for research purposes only. This product
More informationBiochemistry: A Short Course
Tymoczko Berg Stryer Biochemistry: A Short Course Second Edition CHAPTER 10 Carbohydrates 2013 W. H. Freeman and Company Chapter 10 Outline Monosaccharides are aldehydes or ketones that contain two or
More informationGlycan Standards. For microarrays and the identification/ quantification of glycans. Cambridge Isotope Laboratories, Inc. isotope.
Cambridge Isotope Laboratories, Inc. isotope.com RESEARCH PRDUCTS Glycan Standards For microarrays and the identification/ quantification of glycans The emerging field of glycomics focuses on the structure
More informationThe addition of sugar moiety determines the blood group
The addition of sugar moiety determines the blood group Sugars attached to glycoproteins and glycolipids on the surfaces of red blood cells determine the blood group termed A, B, and O. The A and B antigens
More informationContent. Course (LV 21246): Fr. 12:00-14:00 16 Lectures
Content Course (LV 21246): Fr. 12:00-14:00 16 Lectures Content 1: Overview Occurrence of carbohydrates in nature Glycopolymers and glycoconjugates Carbohydrates in animals Carbohydrates in bacteria and
More informationBIOCHEMICAL DIAGNOSIS OF DISEASES CAUSING OLIGOSACCHARIDURIA BY MASS SPECTROMETRY
BIOCHEMICAL DIAGNOSIS OF DISEASES CAUSING OLIGOSACCHARIDURIA BY MASS SPECTROMETRY Pedro Ruiz-Sala, Isaac Ferrer-López Centro de Diagnóstico de Enfermedades Moleculares CEDEM, Universidad Autónoma de Madrid,
More informationMammalian-type Glycosylation l in LEXSY
Mammalian-type Glycosylation l in LEXSY Case Study: Recombinant hu Erythropoietin Jena Bioscience GmbH Loebstedter Str. 80 07749 Jena, Germany Tel.: +49-3641-628-5000 Fax: +49-3641-628-5100 628 e-mail:
More informationDouble charge of 33kD peak A1 A2 B1 B2 M2+ M/z. ABRF Proteomics Research Group - Qualitative Proteomics Study Identifier Number 14146
Abstract The 2008 ABRF Proteomics Research Group Study offers participants the chance to participate in an anonymous study to identify qualitative differences between two protein preparations. We used
More informationIsomer Separation of Positively Labeled N-glycans by CE-ESI-MS
Isomer Separation of Positively Labeled N-glycans by CE-ESI-MS G.S.M. Kammeijer Center for Proteomics and Metabolomics CE IN THE BIOTECHNOLOGY & PHARMACEUTICAL INDUSTRIES 19 TH SYMPOSIUM ON THE PRACTICAL
More informationPr oducts List Ludger Ltd Culham Science Centre Oxford OX14 3EB United Kingdom
Pr oducts List 2018 Ludger Ltd Culham Science Centre Oxford OX14 3EB United Kingdom Email: lily.wang@ludger.com cindy.li@ludger.com www.ludger.com www.ludgersh.com Enzymes for Glycan Release E-PNG01 PNGase
More informationEnzyme Deglycosylation Kit
Contains all enzymes needed to completely remove all N- & simple O-linked carbohydrates from glycoproteins Deglycosylates 2 mg glycoprotein Single reaction at neutral ph Native & denaturing protocols No
More informationMolecular Cell Biology Problem Drill 16: Intracellular Compartment and Protein Sorting
Molecular Cell Biology Problem Drill 16: Intracellular Compartment and Protein Sorting Question No. 1 of 10 Question 1. Which of the following statements about the nucleus is correct? Question #01 A. The
More informationAbdullah zurayqat. Bahaa Najjar. Mamoun Ahram
9 Abdullah zurayqat Bahaa Najjar Mamoun Ahram Polysaccharides Polysaccharides Definition and Structure [Greek poly = many; sacchar = sugar] are complex carbohydrates, composed of 10 to up to several thousand
More informationCharacterization of the Oligosaccharides Associated with the Human Ovarian Tumor Marker CA125*
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 278, No. 31, Issue of August 1, pp. 28619 28634, 2003 2003 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A. Characterization
More informationApplication Note. Abstract. Author. Biotherapeutics & Biosimilars. Sonja Schneider Agilent Technologies, Inc. Waldbronn, Germany
Sensitive and Reproducible Glycan Analysis of Human Immunoglobulin G The Agilent 1260 Infi nity Bio-inert Quaternary LC System with an Agilent AdvanceBio 2.7 µm Glycan Mapping Column and Fluorescence Detection
More informationDetailed Analysis of Polyclonal IgG with Emphasis on Sialylation and Sub-Classes After Fractionation on a Sialic-Acid Binding Lectin
Universität für Bodenkultur Wien Institute of Biochemistry Detailed Analysis of Polyclonal IgG with Emphasis on Sialylation and Sub-Classes After Fractionation on a Sialic-Acid Binding Lectin by Friedrich
More informationExtracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures
Biomolecules 2015, 5, 1741-1761; doi:10.3390/biom5031741 Article OPEN ACCESS biomolecules ISSN 2218-273X www.mdpi.com/journal/biomolecules/ Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific
More informationNature Biotechnology: doi: /nbt Supplementary Figure 1
Supplementary Figure 1 The timeline of the NGAG method for extraction of N-linked glycans and glycosite-containing peptides. The timeline can be changed based on the number of samples. Supplementary Figure
More informationThyroid Function. Thyroglobulin Analyte Information
Thyroid Function Thyroglobulin Analyte Information - 1-2011-01-11 Thyroglobulin Introduction Thyroglobulin (Tg) is a big dimeric protein consisting of two identical subunits. It has 2,748 amino acids in
More informationLudger Guide to Sialylation: II. Highly Sialylated Glycoproteins
Ludger Guide to Sialylation: II Highly Sialylated Glycoproteins Ludger has over 15 years experience providing products and services for the biopharmaceutical industry and in that time we have noticed that
More informationCERTIFICATE OF ANALYSIS
CERTIFICATE OF ANALYSIS PRODUCT NAME: PRODUCT CODE: LOT NUMBER: PACK SIZE: PURITY: FORM: STORAGE: EXPIRATION: GLYKO ASIALO, GALACTOSYLATED, BIANTENNARY COMPLEX N-GLYCAN, CORE-SUBSTITUTED WITH FUCOSE (NA2F)
More informationGlycosaminoglycans: Anionic polysaccharide chains made of repeating disaccharide units
Glycosaminoglycans: Anionic polysaccharide chains made of repeating disaccharide units Glycosaminoglycans present on the animal cell surface and in the extracellular matrix. Glycoseaminoglycans (mucopolysaccharides)
More informationDevelopment of a Glycan Database for Waters ACQUITY UPLC Systems
Mark Hilliard, 1 Weston Struwe, 1 Barbara Adamczyk, 1 Radka Saldova, 1 Ying Qing Yu, 2 John O Rourke, 1 Giorgio Carta, 1 and Pauline Rudd 1 1 National Institute for Bioprocessing Research & Training (NIBRT),
More informationFetuin Glycoprotein Standard
Certificate of Analysis Fetuin Glycoprotein tandard Cat. #: GCP-FET-U-X (GCP-FET-U B7K- *) Batch: BC- Nominal size: * μg Expiry Date: Apr Description: A glycoprotein standard for use during glycan release
More informationon Non-Consensus Protein Motifs Analytical & Formulation Sciences, Amgen. Seattle, WA
N-Linked Glycosylation on Non-Consensus Protein Motifs Alain Balland Analytical & Formulation Sciences, Amgen. Seattle, WA CASSS - Mass Spec 2010 Marina Del Rey, CA. September 8 th, 2010 Outline 2 Consensus
More informationTools for Glycan Analysis
Tools for Glycan Analysis Enzyme Quality & Purity Endoglycosidases QA-Bio enzymes are highly-stable, pure preparations. Enzymes remain active for several days under reaction conditions. QA-Bio enzymes
More informationSupplementary figure 1 Supplementary figure 2
Supplementary figure 1 Schematic overview of the Fc-glycan of IgG. The glycan is composed of a constant core domain (highlighted in red) composed of mannose (Man) and N-acetylglucosamine (GlcNAc) residues
More informationGLYCAN STRUCTURES, CLUES TO THE ORIGIN OF SACCHARIDES
GLYCAN STRUCTURES, CLUES TO THE ORIGIN OF SACCHARIDES Jun Hirabayashi Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University Sagamiko, Kanagawa 199-0195, Japan Tel: 0426-85-3741
More informationAnalysis of N-Linked Glycans from Coagulation Factor IX, Recombinant and Plasma Derived, Using HILIC UPLC/FLR/QTof MS
Analysis of N-Linked Glycans from Coagulation Factor IX, Recombinant and Plasma Derived, Using HILIC UPLC/FLR/QTof MS Ying Qing Yu Waters Corporation, Milford, MA, U.S. A P P L I C AT ION B E N E F I T
More informationCurrent Glycoprotein Analysis. Glycan Characterization: Oligosaccharides. Glycan Analysis: Sample Preparation. Glycan Analysis: Chromatography
Bio Day DENMARK MARCH 2013 Analysis of N-linked Glycans of GlycoProteins marleen_van_wingerden@waters.com Agenda Importance of Glycan Analysis Current Glycoprotein Analysis Glycan Characterization: Oligosaccharides
More information-can be classified by the number of sugars that constitute the molecules: -how to differentiate between glucose and galactose?
Carbohydrates (Also called: saccharides) -can be classified by the number of sugars that constitute the molecules: 1- monosaccharides: -General formula: (CH2O)n -Contain one sugar molecule -Contain two
More informationGlycosaminoglycans, Proteoglycans, and Glycoproteins
Glycosaminoglycans, Proteoglycans, and Glycoproteins Presented by Dr. Mohammad Saadeh The requirements for the Pharmaceutical Biochemistry I Philadelphia University Faculty of pharmacy I. OVERVIEW OF GLYCOSAMINOGLYCANS
More informationIntroduction to Proteomics 1.0
Introduction to Proteomics 1.0 CMSP Workshop Pratik Jagtap Managing Director, CMSP Objectives Why are we here? For participants: Learn basics of MS-based proteomics Learn what s necessary for success using
More informationCell morphology. Cell organelles structure and function. Chapter 1: UNIT 1. Dr. Charushila Rukadikar
UNIT 1 Cell morphology Cell organelles structure and function Chapter 1: Dr. Charushila Rukadikar Assistant Professor Department Of Physiology ZMCH, Dahod Physiology The science that is concerned with
More informationProducts Price List. US Dollars Ludger Ltd Culham Science Centre Oxford OX14 3EB United Kingdom
Products Price List US Dollars 2017 Ludger Ltd Culham Science Centre Oxford OX14 3EB United Kingdom Tel: +44 1865 408 554 Fax: +44 870 163 4620 Email: info@ludger.com www.ludger.com Enzymes for Glycan
More informationThe Automation of Glycopeptide Discovery in High Throughput MS/MS Data
The Automation of Glycopeptide Discovery in High Throughput MS/MS Data Sajani Swamy A thesis presented to the University of Waterloo in fulfilment of the thesis requirement for the degree of Master of
More informationMass Spectrometry. Mass spectrometer MALDI-TOF ESI/MS/MS. Basic components. Ionization source Mass analyzer Detector
Mass Spectrometry MALDI-TOF ESI/MS/MS Mass spectrometer Basic components Ionization source Mass analyzer Detector 1 Principles of Mass Spectrometry Proteins are separated by mass to charge ratio (limit
More informationBiomolecular Mass Spectrometry
Lipids ot different than other organic small molecules Carbohydrates Polymers of monosaccharides linked via glycosidic bonds (acetals/ ketals) many different combinationsvery interesting no time ucleic
More informationRAPID SAMPLE PREPARATION METHODS FOR THE ANALYSIS OF N-LINKED GLYCANS
RAPID SAMPLE PREPARATION METHODS FOR THE ANALYSIS OF N-LINKED GLYCANS Zoltan Szabo, András Guttman, Tomas Rejtar and Barry L. Karger Barnett Institute, Boston, MA, USA PCT Workshop,Boston, 21 May, 2010.
More information5 Identification of Binding Partners of the Annexin A2 / P11 Complex by Chemical Cross-Linking
5 Identification of Binding Partners of the Annexin A2 / P11 Complex by Chemical Cross-Linking In the quest of the omics sciences for holistic schemes, the identification of binding partners of proteins
More informationHelicobacter pylori binding microspheres to prevent gastric cancer
Helicobacter pylori binding microspheres to prevent gastric cancer Inês C. Gonçalves icastro@ineb.up.pt 16 th April 2015 Encontros com a Inovação em Saúde Helicobacter pylori infection World population
More informationCell wall components:
Main differences between plant and animal cells: Plant cells have: cell walls, a large central vacuole, plastids and turgor pressure. The Cell Wall The primary cell wall is capable of rapid expansion during
More informationMain differences between plant and animal cells: Plant cells have: cell walls, a large central vacuole, plastids and turgor pressure.
Main differences between plant and animal cells: Plant cells have: cell walls, a large central vacuole, plastids and turgor pressure. Animal cells have a lysosome (related to vacuole) and centrioles (function
More informationSupporting information
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2014 Supporting information Glycan Reductive Isotope-coded Amino Acid Labeling (GRIAL) for Mass Spectrometry-based
More informationFungal Mannoproteins: the Sweet Path to Immunodominance
Fungal Mannoproteins: the Sweet Path to Immunodominance Mannose residues on glycoproteins trigger key host immune responses that, if better harnessed, could protect against damage and disease Michael K.
More informationElectron transfer dissociation (ETD)
Electron transfer dissociation (ETD) Electron transfer dissociation (ETD) Originally developed for FT-ICR MS as ECD ( C for capture) Now often used on other trapping instruments, even Q-TOFs Trick: Bringing
More informationOligosaccharide Profiling of O-linked Oligosaccharides Labeled with 2 Aminobenzoic Acid (2-AA)
Oligosaccharide Profiling of O-linked Oligosaccharides Labeled with 2 Aminobenzoic Acid (2-AA) Elisabeth A. Kast and Elizabeth A. Higgins GlycoSolutions Corporation, Worcester, MA Data originally presented
More informationGlycoprotein Deglycosylation Kit Cat. No
Visit our interactive pathways at /pathways User Protocol 362280 Rev. 23 February 2006 RFH Page 1 of 5 Glycoprotein Deglycosylation Kit Cat. No. 362280 Note that this user protocol is not lot-specific
More informationIon Source. Mass Analyzer. Detector. intensity. mass/charge
Proteomics Informatics Overview of spectrometry (Week 2) Ion Source Analyzer Detector Peptide Fragmentation Ion Source Analyzer 1 Fragmentation Analyzer 2 Detector b y Liquid Chromatography (LC)-MS/MS
More informationCell Quality Control. Peter Takizawa Department of Cell Biology
Cell Quality Control Peter Takizawa Department of Cell Biology Cellular quality control reduces production of defective proteins. Cells have many quality control systems to ensure that cell does not build
More informationIntroduction. Methods RESEARCH FUND FOR THE CONTROL OF INFECTIOUS DISEASES. TCW Poon *, HLY Chan, HWC Leung, A Lo, RHY Lau, AY Hui, JJY Sung
RESEARCH FUND FOR THE CONTROL OF INFECTIOUS DISEASES Liver specific glycoforms of serum proteins in chronic hepatitis B infection: identification by lectin affinity chromatography and quantitative proteomic
More informationCarbohydrates are aldehyde or ketone compounds with multiple hydroxyl groups Have multiple roles in all forms of life
Carbohydrates 1 Carbohydrates are aldehyde or ketone compounds with multiple hydroxyl groups Have multiple roles in all forms of life Classification Serve as energy stores, fuels, and metabolic intermediates
More informationHCV infects over 170 million people worldwide.
Mass Spectrometric Characterization of Glycosylation of Hepatitis C Virus E2 Envelope Glycoprotein Reveals Extended Microheterogeneity of N-Glycans Roxana E. Iacob, a Irina Perdivara, a,b Michael Przybylski,
More informationProducts Price List. Euros Ludger Ltd Culham Science Centre Oxford OX14 3EB United Kingdom
Products Price List Euros 2018 Ludger Ltd Culham Science Centre Oxford OX14 3EB United Kingdom Tel: +44 1865 408 554 Fax: +44 870 163 4620 Email: info@ludger.com www.ludger.com Enzymes for Glycan Release
More informationDetailed Characterization of Antibody Glycan Structure using the N-Glycan Sequencing Kit
be INSPIRED drive DISCOVERY stay GENUINE APPLICATION NOTE Detailed Characterization of Antibody Glycan Structure using the N-Glycan Sequencing Kit Beth McLeod, New England Biolabs, Inc. Materials Remicade
More informationN-Glycosidase F Deglycosylation Kit
For life science research only. Not for use in diagnostic procedures. FOR IN VITRO USE ONLY. N-Glycosidase F Deglycosylation Kit Kit for the deglycosylation of asparagine-linked glycan chains on glycoproteins.
More informationGlycanPac AXR-1 Columns
CHRMATGRAPHY GlycanPac AXR- Columns For High Resolution Glycan Analysis Product Specifications The Thermo Scientific GlycanPac AXR- columns are highperformance, silica-based HPLC columns for simultaneous
More informationCell Walls, the Extracellular Matrix, and Cell Interactions (part 1)
14 Cell Walls, the Extracellular Matrix, and Cell Interactions (part 1) Introduction Many cells are embedded in an extracellular matrix which is consist of insoluble secreted macromolecules. Cells of bacteria,
More informationStructural and quantitative analysis of N-linked glycans by matrix-assisted laser desorption ionization and negative ion nanospray mass spectrometry
Available online at www.sciencedirect.com Analytical Biochemistry 376 (2008) 44 60 ANALYTICAL BIOCHEMISTRY www.elsevier.com/locate/yabio Structural and quantitative analysis of N-linked glycans by matrix-assisted
More informationendomembrane system internal membranes origins transport of proteins chapter 15 endomembrane system
endo system chapter 15 internal s endo system functions as a coordinated unit divide cytoplasm into distinct compartments controls exocytosis and endocytosis movement of molecules which cannot pass through
More informationTABLE OF CONTENTS GLYCOPRO ENZYMATIC DEGLYCOSYLATION KIT
GLYOPRO ENZYMATI DEGLYOSYLATION KIT ontains all enzymes & reagents needed to completely remove all N-linked & simple O-linked carbohydrates from glycoproteins: Deglycosylates up to 2 mg of glycoprotein
More informationHigher throughput glycosylation analysis of biopharmaceuticals by mass spectrometry
Higher throughput glycosylation analysis of biopharmaceuticals by mass spectrometry Manfred Wuhrer Center for Proteomics and Metabolomics CASSS AT EUROPE, BARCELONA, MARCH 8, 18 GLYCOSYLATION Highly prevalent
More informationGlycoproteomic characterization of recombinant mouse α-dystroglycan
Glycobiology vol. 22 no. 5 pp. 662 675, 2012 doi:10.1093/glycob/cws002 Advance Access publication on January 11, 2012 Glycoproteomic characterization of recombinant mouse α-dystroglycan Rebecca Harrison
More informationStructure & Function of Cells
Anatomy & Physiology 101-805 Unit 4 Structure & Function of Cells Paul Anderson 2011 Anatomy of a Generalised Cell Attached or bound ribosomes Cilia Cytosol Centriole Mitochondrion Rough endoplasmic reticulum
More informationInfluenza A viruses infect the human population on a yearly
Shotgun glycomics of pig lung identifies natural endogenous receptors for influenza viruses Lauren Byrd-Leotis a,1, Renpeng Liu b,c,1,2, Konrad C. Bradley a,3, Yi Lasanajak b,c, Sandra F. Cummings a,c,
More informationCells. Variation and Function of Cells
Cells Variation and Function of Cells Cell Theory states that: 1. All living things are made of cells 2. Cells are the basic unit of structure and function in living things 3. New cells are produced from
More informationIsomeric Separation of Permethylated Glycans by Porous Graphitic Carbon (PGC)-LC-MS/MS at High- Temperatures
Supplementary Information Isomeric Separation of Permethylated Glycans by Porous Graphitic Carbon (PGC)-LC-MS/MS at High- Temperatures Shiyue Zhou 1, Yifan Huang 1, Xue Dong 1, Wenjing Peng 1, Lucas Veillon
More informationJ. Biosci., Vol. 3, Number 4, December 1981, pp Printed in India.
J. Biosci., Vol. 3, Number 4, December 1981, pp. 343-360. Printed in India. Studies on carbohydrate moieties of the glycoprotein, glucoamylase II of Aspergillus niger: Nature of carbohydratepeptide linkage
More informationProfiling the Distribution of N-Glycosylation in Therapeutic Antibodies using the QTRAP 6500 System
Profiling the Distribution of N-Glycosylation in Therapeutic Antibodies using the QTRAP 6500 System Scheduled MRM Pro Algorithm for Increased Efficiency of Targeted Detection Jenny Albanese 1, Christie
More informationStudy of On-Resin Convergent Synthesis of N-Linked. Glycopeptides Containing a Large High Mannose N- Linked Oligosaccharide
Supporting Information Study of On-Resin Convergent Synthesis of N-Linked Glycopeptides Containing a Large High Mannose N- Linked Oligosaccharide Rui Chen and Thomas J. Tolbert* Department of Chemistry,
More informationLudgerPure TM APTS Labelled IgG Glycan Library
Certificate of Analysis LudgerPure TM APTS Labelled IgG Glycan Library Cat. #: CAPTS-IgG-0 Batch #. B-0 Size: approx. 0 pmol Description and: Source A mixture of APTS labelled fucosylated bi-antennary
More informationDiagnosis of CDG Enzyme Analysis and Other Investigations
Diagnosis of CDG Enzyme Analysis and Other Investigations Biochemical Genetics Network Cambridge April 2005 Viki Worthington National Hospital for Neurology and Neurosurgery, London EUROGLYCANET European
More information