Synthetic Oligosaccharides. Indispensable Probes for the Life Sciences

311 Pages·1994·25.17 MB·English

by  Pavol Kováč (Eds.)

ACS SYMPOSIUM SERIES 560
Synthetic Oligosaccharides
Indispensable Probes
for the Life Sciences
1
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0 Pavol Kováč, EDITOR
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6 National Institutes of Health
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August 19, 20 Publication by thoaeft ttDhheiev AiSsiomounetrh oiecfaa sCnt a CRrbheoeghmioynidcaraall t eMS oCecehiteietnmyg,i stry

Johnson City, Tennessee,
October 17-20, 1993
American Chemical Society, Washington, DC 1994
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
Library of Congress Cataloging-in-Publication Data
Synthetic oligosaccharides: indispensable probes for the life sciences /
Pavol Kováč, editor.
p. cm.—(ACS symposium series, ISSN 0097-6156; 560)
"Developed from a symposium sponsored by the Division of
Carbohydrate Chemistry at the Southeast Regional Meeting of the
American Chemical Society, Johnson City, Tennessee, October 17-20,
1993."
Includes bibliographical references and indexes.
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w ISBN 0-8412-2930-9
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56 1. Oligosaccharides—Physiological effect—Congresses. 2. Oligosac­
94-0 cpheaurtiidce us—se—DiCagonnogsrteiscs es.u se—Congresses. 3. Oligosaccharides—Thera­
9
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k- I. Kováč, Pavol, 1938- . II. American Chemical Society. Division
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1/ of Carbohydrate Chemistry. III. American Chemical Society.
02 Southeastern Regional Meeting (1993: Johnson City, Tenn.) IV. Series.
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0.
1 QP702.044S96 1994
s.org 4 | doi: 574.19'24815—dc20 94-168C3I5P
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August 19, 20 Publication AAcChlmolap peRyrtreiiicgrgah hintnts C © tRhh iee1ssm9 e9vrioc4val euldm S. oecT iinehtdeyi c aaptepse atrhaen cceo poyfr igthhet ocwodneer 'as tc othnese nbott tthoamt roefp rtohger afpirhsitc pcaogpeie so fo fe athche

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PRINTED IN THE UNITED STATES OF AMERICA
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
1994 Advisory Board
ACS Symposium Series
M. Joan Comstock, Series Editor
Robert J. Alaimo Douglas R. Lloyd
Procter & Gamble Pharmaceuticals The University of Texas at Austin
Mark Arnold Cynthia A. Maryanoff
University of Iowa R. W. Johnson Pharmaceutical
Research Institute
David Baker
University of Tennessee Julius J. Menn
1 Western Cotton Research Laboratory,
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w0 Arindam Bose U.S. Department of Agriculture
0.f Pfizer Central Research
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5 Roger A. Minear
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4- Robert F. Brady, Jr. University of Illinois
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19 Naval Research Laboratory at Urbana-Champaign
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21/ Margaret A. Cavanaugh Vincent Pecoraro
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0.1 National Science Foundation University of Michigan
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s.org 4 | doi: AUnritvheursri tyB .o fE Wlliissc onsin at Madison MDealmrsohnatl lL aPbhoirlaltioprsie s
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p://pMay Lehigh University North Carolina State University
12 | httDate: Hiroshi Ito A. Truman Schwartz
August 19, 20 Publication MIUBnMaivd eAersllmeitiyan doeef n MP Re.ne Jnsoseyaulrvlclahine iC ae nter MJUonahaicvtna eUl rasRsritbte.ya rS n oChaf- aoIClplllhielnaegomyeis p aign

Lawrence P. Klemann
Nabisco Foods Group L. Somasundaram
DuPont
Gretchen S. Kohl
Dow-Corning Corporation Michael D. Taylor
Parke-Davis Pharmaceutical Research
Bonnie Lawlor
Institute for Scientific Information Peter Willett
University of Sheffield (England)
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
Foreword
IHE ACS SYMPOSIUM SERIES was first published in 1974 to
provide a mechanism for publishing symposia quickly in book
form. The purpose of this series is to publish comprehensive
books developed from symposia, which are usually "snapshots
in time" of the current research being done on a topic, plus
01 some review material on the topic. For this reason, it is neces
0
w sary that the papers be published as quickly as possible.
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6 Before a symposium-based book is put under contract, the
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4-0 proposed table of contents is reviewed for appropriateness to
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9 the topic and for comprehensiveness of the collection. Some
1
bk- papers are excluded at this point, and others are added to
1/
2 round out the scope of the volume. In addition, a draft of each
0
0.1 paper is peer-reviewed prior to final acceptance or rejection.
1
s.org 4 | doi: Terh^is) oanf othney msoyumsp oresivuimew, wphrooc besesc oims es uthpee rveidsietdo r(bsy) othfe t hoer gbaonoikz.
s.ac199 The authors then revise their papers according to the recom
ub5, mendations of both the reviewers and the editors, prepare
p://pMay camera-ready copy, and submit the final papers to the editors,
12 | httDate: who cAhse cak rtuhlaet, aolln nlye coersisgairnya lr erveisseiaorncsh h apvaep ebrese na nmd adoeri.g inal re
August 19, 20 Publication vtiioenws poaf pperresv iaorues liyn pcluubdleisdh eind tphaep evrosl uamree ns.o t Vaecrcbeapttiemd. reproduc
M. Joan Comstock
Series Editor
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
Preface
OLIGOSACCHARIDES ARE COMPONENTS of many important natural
products, and their importance in biological processes has been recog
nized for a long time. In the past, their role could often only be assumed.
Now, the involvement of carbohydrates, and particularly oligosaccharides,
in a variety of vital cellular functions has been proven by solid scientific
evidence. Consequently, activity in the carbohydrate field is constantly
escalating. This situation is documented by the large number of
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0 manuscripts submitted to an increasing number of scientific journals
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0.pr devoted solely to carbohydrates, their conjugates, and their functions.
6
5 Prompted by the progress carbohydrate chemistry has made, particu
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94- larly in the way we can now make synthetic oligosaccharides, together
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k-1 with the ever-increasing importance scientists in the life sciences ascribe
1/b to saccharides, the American Chemical Society decided to organize a sym
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0 posium on synthetic oligosaccharides at its Southeastern Regional Meet
1
10. ing in the fall of 1993 (SERMACS '93). I feel very honored to have been
cs.org 94 | doi: askeWd htoe no rIg asneilzeec ttehda t tmhee ettiintlge., Synthetic Oligosaccharides: Indispensable
p://pubs.aMay 5, 19 Ppinortoseibluleemsc itnuw atilht he a Lnthidfee aSl icfeilea nbsocceireasnt, octerhyse wedxeaesc ricsniiosoetn tamoc ocairsdese oncttihaaatlen. tWhae h ctahet nemtsutaeryr to efda tg hoae s shyaamns
2 | httDate: developed into a branch of carbohydrate chemistry that has given us tools
01n contributing enormously to our better understanding of many fundamen
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19, cati tal processes in the life sciences. The source of most complex oligosac
August Publi cchhaarriiddeess huasveed tirnu lyth bee cliofem es ciinednicsepse niss asbylne thtoeosilss. inT thhuiss ,a rseyan tohfe triecs eoalrigcho.s ac
I was very lucky in the process of organizing the symposium. Some of
the very best carbohydrate chemists active in the preparation and use of
synthetic oligosaccharides accepted my invitation to share some of their
most recent results with the attendees of SERMACS '93. This volume
constitutes a compendium of fine works relevant to the theme of the sym
posium and reflects realistically the state of the art in the discussed topics
as we knew it by the end of 1993.
vii
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
The time frame of SERMACS '93 did not allow me to invite all the
outstanding scientists active in oligosaccharide chemistry and application.
Consequently, this symposium on synthetic oligosaccharides could not
completely cover the subject. To make the permanent record of the sym
posium provided by this volume more comprehensive, I have included a
few invited papers. Of course, no book on such a timely subject can hope
to cover all facets of the topic. Nevertheless, I hope that this volume of
the ACS Symposium Series will be found informative and useful in the
community of researchers and scientists in both academic and industrial
laboratories, and to teachers, students, and all those interested in staying
abreast with the progress in the carbohydrate field.
Acknowledgments
01 The symposium from which this publication originated could not have
0
pr been such a success without the financial assistance obtained from the
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56 Division of Carbohydrate Chemistry of the American Chemical Society, as
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4- well as from the private industry, namely Accurate Chemical & Scientific
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1 Corporation, General Mills, Inc., Glaxo Research Institute, Lederle-Praxis
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b Biologicals, and McNeil Specialty Products Company. The interest of
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02 these organizations in the meeting and their outstanding support are
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0. hereby gratefully acknowledged.
1
cs.org 94 | doi: takeTn heb yp rtohde uActiCoSn Bofo oak ps eDrmeapnaertnmt ernetc. orTd hoef tahses isstyanmcpeo soiuf mo nwea so fu nthdeeirr
a9 members, Anne Wilson, stands out in particular, and I wish to hereby
ubs.5, 1 express my thanks for her efficacious contribution to the project.
p://pMay I am also extremely indebted to Angela Karash for proofreading many
012 | httn Date: hofe ltph, e ams awneulsl craisp ttsh ocuognhtatifnuel dc ionm tmhiesn vtso luonm ei.n dHiveird utaiml eclhya pantedr s cofmropme tetnhte
August 19, 2 Publicatio pPchoAeiVnmOt isLot ,fK hvOaisVe wÁb eČoe nf ntroutl yo innlvy alau alibnleg.u ist but also a very able carbohydrate
National Institutes of Health
Building 8, Room B1A24
Bethesda, MD 20892
Received January 4, 1994
viii
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
Chapter 1
Synthetic Oligosaccharides In Glycobiology
An Overview
Y. C. Lee
Biology Department, Johns Hopkins University,
3400 North Charles Street, Baltimore, MD 21218
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h Glycoconjugates play important roles in biological systems via
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60. recognition of oligosaccharides. Natural glycoconjugates are much
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0 too complicated for the currently available analytical and separation
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99 methodologies to allow dissection of their individual roles in
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10 of definitive structures are very important in understanding
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1 functions of glycoconjugates. Refinement of oligosaccharide
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however, are not as simple or clean-cut as those of nucleic acids or peptides/pr-
oteins. This can be attributed to many factors. Firstly, the chemistiy of
carbohydrates is inherently far more complex than that of other major groups
of biological compounds, even at the basic monomelic level (monosaccharides).
Secondly, assembly of the monomelic units into oligosaccharides raises the level
of complexity. Naturally occurring oligosaccharides can take many closely related
but subtly different forms (glyco-forms, réf. 1). The powerful techniques of
molecular biology have only begun to be effective in elucidating complexity of
carbohydrate structures. Its influence has been mainly in the site-directed
0097-^156/94/0560-0002$08.00/0
© 1994 American Chemical Society
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
1. LEE Synthetic Oligosaccharides In Glycobiology: An Overview 3
mutation to abolish or introduce oligosaccharide chains in glycoproteins or by
modifying the properties of glycosyl-transferases.
Glycoconjugates are those carbohydrates which are covalently linked to
peptides or lipids. The recognition of glycoconjugates in biological systems is
mostly through recognition of the oligosaccharides therein. Often it is only a
small segment of a large polysaccharide that is involved in recognition, as in the
case of heparin. Therefore, to have any clear understanding of the roles of
carbohydrates in glycoconjugates or polysaccharides, one must deal with the
oligosaccharides.
For this purpose, one can isolate and purify oligosaccharides from the
naturally available glycoconjugates involved in biological reactions. This
analytical approach is a conventional but is a veiy difficult approach, requiring
intensive labor and, frequently, expensive instrumentation. Another approach is
01 to synthetically construct oligosaccharides of interest by chemical and/or
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ch enzymatic means. The oligosaccharides to be constructed can be only a partial
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6 representation of the natural carbohydrate structures, or can be artificially
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4-0 modified oligosaccharides substantially different from the natural progenitors.
99 The synthetic approach does not replace the analytical approach, but rather
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k- complements and enhances it. If glycotechnology is to be elevated to a similar
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1 Complexity of naturally occurring glycoconjugates
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glycosyl linkage are possible. When the number of sugar residues increases to
more than three, the situation becomes more complicated because the presence
of many hydroxyl groups on a single sugar allows formation of branched
structures. This is the unique structural feature of carbohydrate as compared
with peptides or nucleotides. When the branching is taking into consideration,
the possible structures that can be generated by a finite number of mono­
saccharides increases exponentially.
Fortunately, natural oligosaccharides do not fully utilize all the possible
structural forms. The number of monosaccharides used in construction of most
of the natural oligosaccharides are quite limited. The sugars used in glycopro­
teins and glycolipids are mostly N-acetyl-D-glucosamine (GlcNAc), N-acetyl-D-
galactosamine (GalNAc), D-mannose (Man), D-galactose (Gal), sialic acids
(SA), L-fucose (Fuc), D-glucose (Glc), and D-xylose (Xyl). For glycosaminogly-
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
4 SYNTHETIC OLIGOSACCHARIDES
cans, D-glucuronic acid (GlcUA), D-iduronic acid (IdoUA), D-glucosamine
(GlcN) are also found. The propensity for certain anomeric forms to be
associated with certain sugars is also quite apparent. GlcNAc is usually found
in β-pyranosyl form including when it is linked to asparagine (Asn) in
glycoproteins, and sialic acids are usually α-linked. In contrast, D-mannose and
D-galactose in natural glycoconjugates can be linked as either a- or β-form. The
β-D-mannopyranosyl linkage occupies an important position in the so-called N-
glycosides of glycoproteins. D-Galactose can be found in nature not only in both
a- and β-anomeric configurations, but also in pyranose and furanose forms (4).
In humans, The α-D-galactopyranosyl residue is a rarity, but it is very common
in rodents.
Neither the overall construction pattern is randoml. There seems to be
only a few "blueprints" that are used for construction of branched oligosacchar­
01 ides of ΛΓ-glycoside in glycoproteins. For example, from the standpoint of RP-
0
h HPLC analyses of pyridylaminated oligosaccharides, the N-glycoside structures
c
60. can be classified into the following four generic patterns (5): M-series
5
4-0 (equivalent to the "mannose-rich" type); X-series (those containing xylose); F-
99 series (those containing fucose); and Z-series (none of the above). This
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k- classification was originally designed for the purpose of analyzing HPLC data
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1/ (parameterization of elution volume) of AMinked oligosaccharide structures, but
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10 the clasification serves to demonstrate limited possibilities of the natural
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1 oligosaccharides in glycoconjugates. Most, if not all of the known "mannose-rich"
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12 | httDate:
August 19, 20 Publication Ma2 Ma 6 1M a 6

Ma2 Ma3
MB 4 Νβ4 NB
Glca3 IIMa2 Ma2 Ma3
Ν : GlcNAc M: Man Glc: Glc
Figure 1. Generic Structure of M-series Oligosaccharides
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.
1. LEE Synthetic Oligosaccharides In Glycobiology: An Overview 5
O-Glycosides often found in the so-called "mucin-type" glycoproteins can
be classified into several groups based on their core structures (6, 7). Unlike N-
glycosides, however, there appears to be no consensus sequence for 0-
glycosylation sites (8), and the glycosylation sites are often veiy close, sometimes
even contiguous.
Although the majority of the N- and O-glycosides seems to be derived
from a limited number of "master plans", many "exceptions" are found quite
frequently, as the analytical instrumentation becomes more sophisticated. For
example, GalNAc rather than GlcNAc is sometimes found to be linked to Asn
in some non-mammammalian glycoproteins (9). Fucose was found to be linked
to Ser/Thr in human Factor IX (10). Glucose-Tyr is found to be the linkage
between glycogen and protein (11). Moreover, there are modifications on the
01 sugar moieties, such as sulfation, phosphoiylation, and methylation. The
0
ch synthetic methodologies utilized must be prepared to cope with these new
60. structural features.
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August 19, 20 Publication GB4 NB4
GB3 Ma 3
Ga3 GB4 NB2
GB4 NB3
Ν : GlcNAc M: Man G: Gal F: L-Fuc
Figure 2. Generic Structure of Z-series Oligosaccharides
In Synthetic Oligosaccharides; Ková, P.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1994.