Table Of ContentPROTEIN GLYCOSYLATION
PROTEIN GLYCOSYLATION
by
Roslyn M. Bili
Gateborg University
Leigh Revers
The Hospital for Sick Children
lain B. H. Wilson
University ofD undee
Foreword by
Harry Schachter
University o/Toronto
lII....
"
SPRINGER SCIENCE+BUSINESS MEDIA, LLC
Library of Congress Cataloging-in-Publication Data
BilI, Roslyn M., 1967-
Protein glycosylation / by Roslyn M. Bill, Leigh Revers, lain B.H.
Wilson; foreword by Harry Schachter.
p. cm.
Includes bibliographical references and index.
ISBN 978-1-4613-7241-7 ISBN 978-1-4615-4939-0 (eBook)
DOI 10.1007/978-1-4615-4939-0
1. Glycoproteins--Synthesis. 1. Revers, Leigh. II. Wilson, lain
B.H. III. Title.
QP552.G59BS5 1998
572' .68-dc21 98-42070
CIP
Copyright © 1998 by Springer Science+Business Media New York
Origina11y published by Kluwer Academic Publishers in 1998
Softcover reprint ofthe hardcover Ist edition 1998
Ali rights reserved. No part ofthis publication may be reproduced, stored in a
retrieval system or transmitted in any form or by any means, mechanical, photo
copying, recording, or otherwise, without the prior written permission of the
publisher, Springer Science+Business Media, LLC.
Printed on acid-free paper.
To Martin, Khamla and Kathi
-WithLove-
Contents
Contributors ix
Abbreviations Xl
Foreword xv
Acknowledgements XIX
SugarsAnd Proteins: GettingItTogether 1
SugarsAnd Proteins: WhyThey GetItTogether 49
Sugars And Proteins: HowThey GetItTogether 85
Core Issues: BuildingTheGroundworkForN-LinkedSugars 147
BranchingOut: ConstructingTheAntennaeOfN-Linked Sugars 213
AndWhatAboutO-LinkedSugars? 281
AddingTheFinishingTouches: TerminalElaborations 329
ExpectThe Unexpected: UnfamiliarGlycosylations 411
ComplementingTheCell: GlycoformSynthesisIn Vitro 457
IcingOnTheCake: SummaryAndFutureDirections 493
Index 499
Contributors
Roslyn M. Bill, M.A.,D.Phil.,M.R.S.C.,C.Chem.
TheLundbergLaboratory, UniversityofGoteborg,
Box462, S40530, GOteborg, Sweden.
LeighRevers, M.A.,D.Phil.
DepartmentofBiochemistryResearch, TheHospitalforSickChildren,
555UniversityAvenue,Toronto, Ontario, M5G IX8, Canada.
lainB. H. Wilson, B.Sc.(Hons),D.Phil.,MRS.C.,C.Chem.
DepartmentofBiochemistry, The WellcomeTrustBuilding, UniversityofDundee,
DowStreet, Dundee, DDI5EH, Scotland.
Abbreviations
The symbols x and Xrepresent unspecified Arabic and Roman numerals,
respectively.
Ac Acetyl.
AGE Advancedglycationend-product.
Amb y-Aminobutyrine(anaminoacid).
Ara! L-Arabinofuranose.
Bz Benzyl.
CD 'Clusterofdifferentiation' leukocyteantigen,
e.g. CD2, CD43 (also known as leukosialin),
etc.
cDNA ComplementaryDNA.
CHO Chinesehamsterovary.
CMP Cytidine5'-monophosphate.
COS Acell-linederivedfromAfrican green monkey
kidney cells (CV-l) by transformation with
theorigin-defective simian virus 40(SV40);
commonlyused in mammaliantransient
transfectionexperiments.
CR Complementreceptor.
CRD Carbohydraterecognitiondomain.
CSL Cerebellarsoluble lectin.
DCCI Dicyclohexylcarbodiimide.
DEPC Diethylpyrocarbonate.
DNA 2'-Deoxyribonucleicacid.
Dol Dolichol,dolichyl.
Dol-P-Glc Dolichyl phospho-~-D-glucose.
xii Abbreviations
Dol-P-Man Dolichylphospho-~-o-mannose.
ECM Extracellularmatrix.
ECa.b.c.d UniqueenzymedesignationoftheEnzyme
Commission ofthe International Union of
Biochemistry(a, b, c anddare Arabic
numerals).
EDTA Ethylenediaminetetraaceticacid.
EEDQ N-Ethoxycarbonyl-2-ethoxy-I,2-
dihydroquinoline.
EGF Epidermal growthfactor.
ER Endoplasmic reticulum.
ESL-I E-Selectinligand-I.
EST Expressed sequencetag.
Fmoc Fluoren-9-yl-methoxycarbonyl.
Fuc L-Fucose.
Fuc-TX Various human al,3-fucosyltransferases
numberedaccordingtothedateofcloningof
thecorrespondingDNA.
GAG Glycosaminoglycan.
Gal o-Galactose.
Galf o-Galactofuranose.
GalNAc N-Acetyl-o-galactosamine.
GalNAc-Tx Various UDP-GaINAc:polypeptideGaINAc
transferases numberedaccordingtothedate
ofcloningofthecorrespondingDNA.
GDP Guanosine-5'-diphosphate.
GIPL Glycoinositolphospholipid.
Gle o-Glucose.
GleNAc N-Acetyl-o-glucosarnine.
GleNAc-TX Varioushuman ~-N-acetylglucosaminyl
transferases numbered accordingtothe
methodofSchachter.
GlyCAM-1 Glycosylation-dependentcell-adhesion
molecule-I.
GPI Glycosylphosphatidylinositol.
GPT GleNAc-I-Phosphotransferase.
HBV HepatitisB virus.
hCG Humanchorionic gonadotropin.
HEV Highendothelial venules.
hGM-CSF Human granulocyte-macrophagecolony
stimulatingfactor.
HN Human immunodeficiency virus.
Abbreviations xiii
hLF Human lactoferrin.
HOBt N-Hydroxybenzotriazole.
HPAEC High performance anion-exchange
chromatography.
IgG Immunoglobulin G.
Kdo 2-Keto-3-deoxynonulosonic acid.
K TheMichaelisconstantofenzymekinetics.
m
Lecx Oneofa seriesoflectin-resistantCHOcell-lines
exhibitinga recessivecharacter.
LECx One ofaseries oflectin-resistantCHOcell-lines
exhibitingadominantcharacter.
Lea, Leb, LeX, LeY Lewis bloodgroupand relatedantigens.
LPG Lipophosphoglycan.
MAdCAM-I Mucosaladressincell-adhesion molecule-I.
Man o-Mannose.
ManNAc N-Acetyl-o-mannosamine.
MOCK Madin-Oarbycaninekidney.
Me Methyl.
M Relative molecular mass.
r
mRNA MessengerRNA.
NBuONJ N-Butyldeoxynojirimycin.
NCAM Neuralcell-adhesion molecule.
Neu5Ac N-Acetylneuraminic acid.
Neu5Gc N-Glycolylneuraminicacid.
NMR Nuclearmagnetic resonance.
OGT UDP-GlcNAc:polypeptideGlcNAc-transferase
(O-GlcNAc transferase).
OPfp Pentafluorophenylester.
Om Ornithine(anaminoacid).
p
Inorganic phosphate.
PAGE Polyacrylamidegelelectrophoresis.
PAPS 3'-Phosphoadenosine-5'-phosphosulphate(the
sulphotransferasedonor).
PCR Polymerasechainreaction.
Pd(o) Palladiummetal.
PNPase Nucleoside phosphorylase.
pNP-Man p-Nitrophenyl-a-mannoside.
pp Inorganic pyrophosphate.
ppGalNAc-transferase UOP-GaINAc:polypeptideGaINAc-transferase.
PSGL-I P-selectin glycoproteinligand-I.
PUGNAc O-(2-Acetamido-2-deoxy-o-gluco-
pyranosylidene)amino-N-phenyl-carbamate.
xiv Abbreviations
Qui3NAc 3-N-Acetylquinovosamine.
RA Rheumatoidarthritis.
Rha Rhamnose.
RNA Ribonucleic acid.
RNase Ribonuclease.
SDS Sodiumdodecyl sulphonate.
Sia Sialicacid.
sialyl-Lea Sialyl-Lewisaantigen.
sialyl-Lex Sialyl-Lewisxantigen.
S-layer Surfacelayer.
SRP Signal recognition particle.
ST3GalX Various a2,3-sialyltransferasestransferringto
galactoseresidues.
ST6GalX Various a2,6-sialyltransferasestransferringto
galactoseresidues.
ST6GalNAcX Variousa2,6-sialyltransferasestransferring to
GalNAc residues.
ST8SiaX Various a2,8-sialyltransferasestransferringto
sialic acid residues.
TAP24 ApeptidederivedfromtheMUCI mucin,
specifically: Thr-Ala-Pro-Pro-Ala-His-Gly
Val-Thr-Ser-Ala-Pro-Asp-Thr-Arg-Pro-Ala
Pro-Gly-Ser-Thr-Ala-Pro-Pro.
TAP25 A peptidederivedfromthe MUCI mucin,
specifically: Thr-Ala-Pro-Pro-Ala-His-Gly
Val-Thr-Ser-Ala-Pro-Asp-Thr-Arg-Pro-Ala
Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala.
t-Bu Tertiary butyl.
THF Tetrahydrofuran.
tPA Tissue plasminogenactivator.
TRG Tunicamycin resistance gene.
Tyv Tyvelose, 6-dideoxY-D-arabino-hexopyranose.
UDP Uridine-5'-diphosphate.
V Limitingreaction velocity.
rnax
VSG Variantsurfaceglycoprotein.
VSV Vesicularstomatitis virus.
Xyl D-Xylose.
XylMU 4-Methylumbelliferyl-p-xyloside.
