REvIEWS

Glycointeractions in bacterial
pathogenesis
Jessica Poole1, Christopher J. Day1, Mark von Itzstein   1, James C. Paton2 and
Michael P. Jennings   1*
Abstract | Many important interactions between bacterial pathogens and their hosts are highly
specific binding events that involve host or pathogen carbohydrate structures (glycans). Glycan
interactions can mediate adhesion, invasion and immune evasion and can act as receptors for
toxins. Several bacterial pathogens can also enzymatically alter host glycans to reveal binding
targets, degrade the host cell glycans or alter the function of host glycoproteins. In recent years,
high-​throughput screening technologies, such as lectin, glycan and mucin microarrays, have
transformed the field by identifying new bacterial–host glycointeractions, which are crucial for
colonization, persistence and disease. In this Review , we discuss interactions involving both host
and bacterial glycans that have a role in bacterial pathogenesis. We also highlight recent
technological advances that have illuminated the glycoscience of microbial pathogenesis.

Lectin
A carbohydrate-​binding protein
that is not an anti-​carbohydrate
antibody and does not modify
the carbohydrate.
Glycan
A generic term for any sugar or
assembly of sugars in free form
or attached to another
molecule that is
interchangeable with the terms
saccharide and carbohydrate.
Mucin
A large, heavily glycosylated
protein that is the main
component of mucus.
Glycome
Carbohydrates present in a
biological sample.
1
Institute for Glycomics,
Griffith University, Gold
Coast Campus, Gold Coast,
Australia.
2
Research Centre for
Infectious Diseases,
Department of Molecular
and Biomedical Science,
University of Adelaide,
Adelaide, Australia.
*e-​mail: m.jennings@
griffith.edu.au
https://doi.org/10.1038/
s41579-018-0007-2
Although carbohydrates are the most abundant organic
molecules in nature1 and are found in every domain of
life2, identifying their biological roles has been difficult.
However, in the past decade, high-​throughput meth-
ods, such as lectin3–5, glycan6,7 and mucin microarrays8,
have allowed the ‘glycome’ to be deciphered — a devel-
opment that resembles the expansion of knowledge of
the genome and proteome that other high-​throughput
methods have provided.
Pathogenic bacteria present glycosylated mole-
cules to their host, such as capsular polysaccharides,
glycoproteins and lipooligosaccharides (LOS) or
lipopolysaccharides (LPS), which can have pivotal
roles in colonization and disease 9,10. Glycans on the
surface of host cells are used by many pathogens for
attachment and invasion11–15, as a carbon source16,17
and as the targets of bacterial toxins18–20. Furthermore,
bacterial glycosyltransferases and glycosidases can
modify host glycans as part of the pathogenic pro-
cess. The study of glycointeractomics explores these
host–pathogen interactions that utilize either host-​
associated or pathogen-​a ssociated glycans. In this
Review, we discuss interactions involving both host
and bacterial glycans that have a role in bacterial
pathogenesis. We also highlight recent technological
advances that have illuminated the glycoscience of
microbial pathogenesis.
Bacterial adhesins and host glycans
Many bacterial lectins act as adhesins11–15,21, which medi-
ate attachment to host cells, initiate colonization and
define bacterial cell and tissue tropism.
Fimbrial adhesins FimH and UclD of Escherichia coli.
Escherichia coli is an important human pathogen that can
cause a range of diseases from enteritis to urinary tract
infections (UTIs) and meningitis; for example, uropatho­
genic E. coli (UPEC) is a major cause of UTIs22,23. E. coli
uses several different types of pili to adhere to host sur-
faces (for example, fimbrial adhesin PapG24, type 1 fim-
brin d-​mannose specific adhesin (FimH)15 and UclD21),
and the binding specificities of these pili determine
host and tissue tropism. Type 1 pili carry the lectin FimH
at their tip, which mediates binding to mannose on the
surface of host cells. FimH from E. coli that has colo-
nized the large intestine of healthy humans preferentially
binds monomannosylated host glycans, whereas FimH
from E. coli isolated from UTIs preferentially binds
oligomannose host glycans 15 (Fig.  1a) . This prefer-
ence for oligomannose by UPEC has been used as
the basis for rational drug design of C1-modified
α-​mannoside-based FimH antagonists. A class of manno-
sides, biphenyl α-d-mannopyranosides, has been tested in
mouse models and has shown potential as UTI therapeu-
tics25–28. It has recently been reported that another E. coli
pilus adhesin, UclD, has lectin function. The UclD lectin
domain binds to colonic epithelial cells, and this inter-
action was inhibited by pretreatment of O-​glycosidase
to remove O-​linked glycans, suggesting UclD interacts
with one or more O-​linked glycans on host cells21.
The BabA, SabA and LabA adhesins of Helicobacter
pylori. The Gram-​negative bacterium Helicobacter
pylori colonizes the gastric mucosa of over half the
global human population29. H. pylori causes a persistent

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Pathogen Host
a

FimA
FimD FimH
E. coli R Uroepithelial cell
FimF FimG α3 α6

b BabA LeB β3
R
α2 α4

SabA sLeX β4
R
Gastric epithelial cell
H. pylori α3 α3

LabA
β4
R MUC5AC
LacdiNAc

c M protein

GAS R Oral epithelial cell

β3 β3 β4
α2
Blood group H
(type 1)

d
α3
CR3

I-domain Cervical
N. gonorrhoeae epithelial cell
Type IV pilin

Gal GalNAc Fuc diNAcBac Man

Glc GlcNAc Neu5Ac GlcA R = any underlying linkage

Fig. 1 | Adherence by pathogenic bacteria mediated by bacterial or host lectins. a | Type 1 fimbrin d-​mannose
specific adhesin FimH of Escherichia coli targets oligomannose that is expressed on the host uroepithelium during
urinary tract infections. b | BabA , SabA and LabA are adhesins of Helicobacter pylori. BabA interacts with Lewis B (LeB)
expressed on gastric epithelial cells. SabA binds the glycans sialyl Lewis X (sLeX) and sialyl Lewis A. These sialylated
structures are expressed on the host gastric lining during a chronic infection. LabA interacts with a lacdiNAc structure
(GalNAcβ1-4GlcNAc) present on the mucin MUC5AC in the gastric mucosal layer. c | Human oral epithelial cells are
glycosylated with ABO blood group antigens. The M protein of group A Streptococcus (GAS) interacts with these
antigens with the highest affinity for the blood group H antigen. d | The pili of Neisseria gonorrhoeae can be glycosylated
with the disaccharide Gal(α1-3)diNAcBac when the glycosyltransferase gene pglA is expressed. This disaccharide interacts
with the I-​domain of complement receptor 3 (CR3) present on cervical epithelial cells and allows for successful gonococcal
attachment and invasion. BabA , blood group antigen-​binding adhesin; diNAcBac, N,N′-diacetylbacillosamine; Fuc, fucose;
Gal, galctose; GalNAc, N-​acetylgalactosamine; Glc, glucose; GlcA , glucuronic acid; GlcNAc, N-​acetylglucosamine; LabA ,
lacdiNAc-​binding adhesin; Man, mannose; Neu5Ac, N-​acetylneuraminic acid; SabA, sialic acid-binding adhesin.

Glycosyltransferases
Enzymes that catalyse the
transfer of a sugar from a sugar
nucleotide donor to a
substrate.
infection with a majority of hosts being asymptomatic30.
However, 1–3% of those infected develop gastric can-
cer, and as such, H. pylori is considered a carcinogen
by the WHO30,31. The human gastric mucosa and epi-
thelial lining are heavily glycosylated, and H. pylori uses
two lectins to bind the glycosylated structures within
this environment — blood group antigen-​binding
adhesin BabA and sialic acid (also known as neu-
raminic acid)-binding adhesin SabA. BabA is specific
for fucosylated glycoconjugates such as Lewis B, which is

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Glycosidases
Enzymes that catalyse the
hydrolysis of the bonds
between two saccharides in a
glycan.
Glycointeractomics
The systematic study of
interactions between biological
entities mediated by glycans.
Adhesins
A protein on the surface of
bacteria, viruses or parasites
that binds to a ligand present
on the surface of a host cell.
Glycoconjugates
Molecules in which one or more
glycan units are covalently
linked to a non-​carbohydrate
entity.
Lewis B
Fucα1-2Galβ1-3(Fucα1-4)
GlcNAc, a tetrasaccharide found
as a terminal modification on
both O-​linked and N-​linked
glycans.
Lewis blood group antigens
A related set of glycans that
carry a non-​teminal α1-3 or
α1-4 fucose residues covalently
linked to N-​acetylglucosamine.
Sialyl Lewis X
(sLeX). Neu5Acα2-3Galβ1-
4(Fucα1-3)GlcNAc, a
tetrasaccharide found as a
terminal modification on both
O-​linked and N-​linked glycans.
Sialyl Lewis A
Neu5Acα2-3Galβ1-3(Fucα1-4)
GlcNAc, a tetrasaccharide found
as a terminal modification on
both O-​linked and N-​linked
glycans.
LacdiNAc
GalNAcβ1-4GlcNAc, a
disaccharide found as a
terminal modification on both
O-​linked and N-​linked glycans.
MUC5AC
A mucin encoded by the
MUC5AC gene.
Group A Streptococcus
(GAS). Streptococcus belonging
to the Lancefield grouping
based on a serological
methodology for classifying
Streptococcus species;
Streptococcus pyogenes is
the species defined by group
A Streptococcus.
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expressed by gastric epithelial cells and found on mucins dependent on the sex of the host 40. The terminal
within the mucosa12 (Fig. 1b). The adherence that is medi- lacto-​N -neotetraose structure that is expressed on
ated by BabA is pH dependent32. BabA binding is most N. gonorrhoeae strain 1291 LOS binds to asialoglyco-
efficient in the higher pH environment of the gastric protein receptors (ASGRs), mediating the adherence
mucosa. SabA is specific for the Lewis blood group antigens and invasion of male primary urethral epithelial cells41.
sialyl Lewis X (sLeX) and sialyl Lewis A13 (Fig. 1b). Low lev-
The gonococcus also expresses type IV pili42, glyco-
els of sialic acids are expressed in the gastric lining of protein adhesins that are crucial for the establishment
healthy individuals. Sialic acid expression is increased of infection in females. N. gonorrhoeae strain 1291 pili
in the stomach lining after H. pylori has established a are glycosylated with either a monosaccharide N,Nʹ-
chronic infection31, demonstrating that H. pylori influ- diacetylbacillosamine (diNAcBac) or a disaccharide
ences the host gastric glycome. The majority of H. pylori Gal(α1-3)diNAcBac by glycosyltransferases encoded
colonizes the gastric mucosa, but only ~20% of the bac- by pilin glycosylation (pgl) genes43, depending on the
terial population is directly attached to the gastric epi- phase variable PglA glycosyltransferase43–47. When pglA
thelium33, suggesting that the infected mucous layer acts is expressed, the pili are glycosylated with the disac-
as a reservoir for persistent infection of the gastric epi- charide, and they are able to interact with the I-​domain
thelium. Recently, another adhesin, LabA (also known of the α-​subunit of complement receptor 3 (CR3; also
as lacdiNAc-​binding adhesin) of H. pylori was observed known as ITGAM/ITGB2, Mac-1 and CD11b/CD18),
to have glycan-​binding activity. LabA was found to bind which is present on human primary cervical epithelial
lacdiNAc structures (GalNAcβ1-4GlcNAc) conjugated cells48,49 (Fig. 1d). This finding was the first evidence that
to MUC5AC mucins in the gastric mucosal layer14 (Fig. 1b). the I-​domain of CD11b has lectin activity48. The interac-
This adhesin may have a role in persistence by retain- tion between the disaccharide and the CD11b I-​domain
ing H. pylori in the mucosal layer and not always being allows the attachment and invasion of the cervical epithe-
attached to the stomach epithelial cells. lia by the gonococcus. If this disaccharide is absent, the
gonococcus is hyperinvasive to cervical epithelial cells
M protein of Streptococcus pyogenes. Streptococcus but is unable to survive within the cells48. In summary,
pyogenes (group A Streptococcus (GAS)) is a Gram-​ Gal(α1-3)diNAcBac glycosylation of gonococcal pilin
positive bacterium that causes a range of human dis- and its interaction with the host lectin CR3 facilitates the
eases, including acute pharyngitis and skin diseases such colonization and invasion of host cervical cells.
as impetigo. Pharyngeal infection with GAS can develop
into rheumatic fever, a leading cause of heart disease for Immune evasion
children in developing nations34. M protein — a major The ability of bacterial pathogens to colonize and per-
virulence factor of GAS and a trigger of the autoimmune sist within a host depends on successful evasion of host
reaction that leads to rheumatic fever34 — interacts with immune responses. There are a range of mechanisms
glycosaminoglycans for adherence to host cells35. Strains whereby glycointeractions mediate evasion of the host
that belong to the M1T1 GAS globally dominant, highly immune system. For example, bacteria produce surface
virulent clone36 are frequently isolated from throat cul- structures, such as capsules, to inhibit the host immune
tures. A recent study demonstrated that M protein of functions. Some bacteria produce structures that mimic
M1T1 binds ABO(H) blood group antigens , which are host glycan structures to subvert host recognition. Box 2
highly expressed on oral epithelial cells and in mucosal provides examples of bacterial glycans that mimic host
fluid11,37, but had a preference for H antigen over A structures and how this may promote autoimmunity. Also,
and B antigens11 (Fig. 1c). The ability of M protein to bind some pathogens can express glycans on proteins or other
certain classes of glycans may have a major role in host surface molecules that inhibit host immune functions.
tissue tropism and disease outcome.
Capsular polysaccharides. Bacterial capsular polysac-
Host lectins involved in bacterial adhesion charide structures that mimic host glycan structures are
Host lectins often have roles in the immune system a common mechanism of immune system evasion. GAS
and have evolved to initiate the host immune response expresses a capsule comprising hyaluronan (HA)10,50,
to glycans produced by multiple pathogens, such as the which is the most abundant glycosaminoglycan in the
C-​type lectin dendritic cell-​specific ICAM-3-grabbing skin51. The length of the chains of HA influences the sur-
non-​integrin 1 (DC-​SIGN; also known as CD209)38 and vival of GAS in skin and deep tissue infections through
the mannose-​binding lectin39. Pathogenic bacteria that interactions with the cell surface receptor CD44 (ref.51).
interact with host lectins have to display structures Interactions between CD44 and HA are involved in
that enable bacteria to adhere to and/or to enter the cell lymphocyte recruitment during inflammation52. A GAS
without triggering the lectin response to bacteria. Recent capsule comprising low molecular weight HA causes
work has described numerous interactions between greater uptake of the bacteria by macrophages (Fig. 2a),
bacterial and host glycans (Box 1). whereas uptake of GAS with a high molecular weight
HA capsule is lower. A high molecular weight HA cap-
Glycosylated pili of Neisseria gonorrhoeae and sule also confers higher GAS survival rates in blood kill-
complement receptor 3. Neisseria gonorrhoeae (the ing assays. This effect of low and high molecular weight
gonococcus) is an obligate human pathogen that capsular HA on the survival of GAS depends on the cell
uses two different host lectins to mediate adherence; surface receptor CD44, as CD44 knockout mice do not
the host lectin that is utilized by the gonococcus is show a difference in response to HA molecular weight
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Glycosaminoglycans
in the GAS capsule51. The observation that the molecular Binding of sialylated structures to ITIM mediates down-
Long unbranched weight of HA attached to the GAS capsular surface can regulation of signalling pathways, which helps maintain
polysaccharides consisting of a influence uptake by macrophages suggests that glycans a baseline state in innate immune cells and contributes to
repeating disaccharide unit on the bacterial surface assist in immune system evasion. anti-​inflammatory responses56. All group B Streptococcus
composed of a hexose
derivative and an amino sugar
Siglecs are immunoglobulin-​like lectins that bind (GBS) serotypes express terminal α2-3-linked N-​
derivative that forms the sugar sialic acids and have roles in both the innate and adap- acetylneuraminic acid (Neu5Ac) capsular polysaccha-
portion of a proteoglycan. tive immune system53,54. CD33-related Siglecs are a subset rides — the predominant sialic acid that is expressed on
of Siglecs that often contain an immunoreceptor tyrosine-​ human cells57,58. The N-​acetylneuraminic acid on GBS
based inhibitory motif (ITIM) and an ITIM-​like motif55. capsules is bound by CD33-related Siglec-9 and Siglec-5
on human leukocytes58. The carbohydrate Siaα2-3Galβ1-
4GlcNAc (a polysaccharide expressed on the GBS cap-
Box 1 | Glycan–glycan interactions sule in serotypes Ia, Ib, III and V (ref.58)) interacts with
Glycan–glycan interactions were once assumed to be weak, low-​affinity binding events Siglec-9 (ref.9) (Fig. 2b). The engagement of this sialylated
between two interacting partners that preceded the more important protein–glycan capsule carbohydrate with Siglec-9 dampens neutrophil
interactions137–139. the first glycan–glycan interaction to be recorded was in sea responses, such as oxidative bursts and the formation of
sponges. sea sponges can be homogenized into single cells and reform into whole neutrophil extracellular traps9. GAS engages a different and
organisms; this process was found to be species-​specific and dependent on glycan– as yet undefined binding site on Siglec-9, which binds HA
glycan interactions between proteoglycans found on the sponge cells of the same in the GAS capsule10 (Fig. 2b). This is the first example of
species140–142. Later, glycan–glycan interactions were reported in mammals; roles were a CD33-related Siglec binding a non-​sialylated carbohy-
described for Lewis X and ganglioside homodimers in mouse embryo development143
drate10. This shows that GBS and GAS have both evolved
and cancer144,145, respectively. it has recently been reported that direct interactions
between bacterial glycans and host glycans can mediate host–pathogen
to use the host lectin Siglec-9 for immune evasion.
interactions116,117. the glycolipid diglucosyldiacylglycerol of Enterococcus faecalis has Neisseria meningitidis is a human-​adapted pathogen
been shown to interact with the glycosaminoglycans heparin and heparan sulfate on and is a leading cause of septicaemia and bacterial men-
the surface of host cells146. the glycan portion of lipooligosaccharide (LOs) or ingitis. N. meningitidis is a commensal in the nasophar-
lipopolysaccharide (LPs) of Campylobacter jejuni, Neisseria meningitidis, Salmonella ynx of approximately 5–10% of healthy humans59,60. The
enterica subsp. enterica serovar typhimurium, nontypeable Haemophilus influenzae N. meningitidis serogroup C conjugate (MCC) vaccine
(NtHi) and Shigella flexneri has also been shown to interact directly with host cell consists of α2-9-linked polysialic capsular polysaccharide
surface glycans, and this binding was observed to be important for adherence to conjugated to a carrier protein61,62. Protection is achieved
epithelial cells in culture116,117. several of these species (C. jejuni, N. meningitidis and by the generation of bactericidal antibodies that bind to
NtHi) exhibit host molecular mimicry, making the terminal structures of these LOs or
the serogroup C capsule62–64. Three N. meningitidis sero-
LPs glycans identical to host glycan structures. the highest-​affinity glycan–glycan
interactions that have been identified so far have been between host glycoconjugates
group C vaccine escape strains have been reported that
and bacterial LOs or LPs that mimic host glycans. For example, the asialo-​GM1 terminal display increased resistance to the MCC vaccine bacte-
structure made by C. jejuni interacts with the host glycan, blood group B antigen ricidal antibodies62. The resistance correlated with the
(see the figure, part a), with a dissociation constant of 140 nM (ref.116) and 3′-sialyl-​ presence of an additional insertion sequence (IS), IS1301,
lacto-N-​neotetraose made by N. meningitidis interacting with thomsen–Friedenreich in the intergenic region between the sia and ctr operons
(tF) tumour antigen on the host cell surface (see the figure, part b), with a dissociation that are required for the biosynthesis and export of cap-
constant of 13 nM (ref.117). the observation that host-​mimic structures allow for such sule. This additional IS causes an increase in transcript
high-​affinity interactions indicates a much broader and yet to be understood role for levels for both operons, resulting in the synthesis of more
glycan–glycan interactions beyond host–pathogen adherence. capsular polysaccharide62. Complement activation is
a b N. meningiditis vital for immunity to N. meningitidis65, and, paradoxi-
C. jejuni
cally, although serotype C capsular polysaccharide is the
target antigen for the protective antibodies, the increase
in capsule polysaccharide is associated with impairment
R
R of complement activation by the alternative pathway62
β4 (Fig. 2c). Capsules that contain sialic acid can interfere
with the amplification of the complement cascade by
β4 β4 inhibiting the alternative pathway of complement acti-
vation, which results in a decrease in C3 and membrane
β3 β3
Blood group B attack complex deposition on the bacterial surface62,66–68.
(type V) α3 TF antigen
Asiolo-GM1
β4 Bacterial protein glycosylation. Both O-​linked and
α2 -Sialyl lacto-N
β4 β3 neotetraose N-​linked glycosylation systems have been identified in
α3 pathogenic bacteria, with O-​linked glycosylation pathways
more common than N-​linked69. O-​linked glycosylation
R R was first identified in Neisseria species70,71, with the first
peptide glycosyltransferase identified in N. meningitidis43.
Epithelial cell
Campylobacter jejuni, a commensal organism of chickens
Gal GalNAc Fuc Neu5Ac
and a food-​borne pathogen in humans, was the first bac-
Glc GlcNAc
terium in which an N-​linked protein glycosylation system
R = any underlying linkage
was identified72,73. This ubiquitous pathway is now known
Fuc, fucose; Gal, galactose; Glc, glucose, GalNac, N-acetylgalactosamine; GlcNac, to post-​translationally modify over 60 C. jejuni pro-
N-acetylglucosamine; Neu5ac, N-acetylneuraminic acid. teins74. It has been observed that C. jejuni with mutations

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ABO(H) blood group
antigens
Carbohydrate structures
attached to cells and secreted
proteins that determine the
blood group and/or type of a
person.
Phase variable
A method of responding to
varying environments that
allows the switching of gene
expression in an on–off
fashion.
Immunoreceptor tyrosine-​
based inhibitory motif
(ITIM). A conserved sequence
of amino acids found in the
cytoplasmic tails of many
inhibitory receptors of the
immune system.
Group B Streptococcus
(GBS). Streptococcus belonging
to the Lancefield grouping
based on a serological
methodology for classifying
Streptococcus species;
Streptococcus agalactiae is the
species defined by group B
Streptococcus.
Oxidative bursts
Release of reactive oxygen
species; also known as
respiratory bursts.
Neutrophil extracellular
traps
Networks of fibres, primarily
composed of DNA, used to
bind pathogens.
Insertion sequence
(IS). A short (<2500 base pairs)
transposable segment of DNA.
N-​linked glycosylation
Covalent attachment of a
carbohydrate through a bond
to the amide nitrogen of an
asparagine residue.
O-​linked glycosylation
Covalent attachment of a
carbohydrate through a bond
to the hydroxyl group of a
serine or threonine residue.
Nontypeable Haemophilus
influenzae
Unencapsulated strains of
H. influenzae.
Toll-​like receptor-5
A membrane-​spanning innate
immune system protein known
to detect bacterial flagellin.
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Box 2 | Molecular mimicry and autoimmune disease
Many bacteria have been identified that produce surface glycosylations that mimic host structures, including Neisseria
meningitidis serogroup B capsule147, group a Streptococcus (Gas)10,50 and group B Streptococcus (GBs) capsules, and the
lipooligosaccharide (LOs) of pathogenic Neisseria spp., Campylobacter jejuni, Helicobacter pylori and Haemophilus
influenzae83,148. Molecular mimicry of host glycans carries a risk of induction of self-​antibody production.
For example, N. meningitidis serogroup B was not included in the broad-​coverage glycoconjugate vaccine that is used for
N. meningitidis owing to the similarity between the serogroup B capsule (repeating α2-8Neu5ac) and host sialylglycocon-
jugates147. there are numerous cases of pathogen-​induced autoimmune diseases149–151. However, relatively few of these
have been directly correlated with the glycosylation profile of the bacteria83. the best-​studied example of a pathogen-​
induced autoimmune disease is Guillain–Barré syndrome. this disease causes ascending neuromuscular paralysis. the
C. jejuni LOs mimics the carbohydrate GM1 ganglioside that is expressed on host cells148,152 (see the figure). a variant form
of Guillain–Barré syndrome known as Fisher syndrome has also been correlated to ganglioside mimicry in C. jejuni and
sialyllactose mimicry in nontypeable Haemophilus influenzae (NtHi) and type b H. influenzae83. it has long been noted that
peptides can be used to elicit immune responses that are equivalent to a complex glycan, making it possible to make
peptide vaccines for complex, difficult-​to-synthesize glycans153. a recent study has demonstrated that the process that
leads to autoimmunity can work in reverse, with bacterial glycans that mimic host peptides eliciting an immune response
that leads to anti-​host-protein antibody production. this discovery has led to an understanding of the molecular basis of
some autoimmune diseases, such as autoimmune thyroid diseases154. For example, linear α-1-6 glucans that are produced
by the probiotic bacterium Bifidobacterium bifidum are
reported to be cross-​reactive with human serum thyroid
peroxidase and thyroglobulin autoantibodies, providing Guillain–Barré
β3 syndrome GM1
evidence of a potential role of α-1-6 glucan that is produced
by bacteria in generating autoimmune thyroid diseases154. β3
the study of peptide-​glycan or glycan-​peptide mimicry in β4
α3
the context of autoimmunity is an emerging area, indicating β4
that further roles for glycans of bacterial origin may be β4 α3
Molecular
discovered in autoimmune diseases. research on the gut mimicry β4
microbiome has shown that gut commensals also have a vital
role in the host immune system through glycointeractions. R
For example, the human intestinal commensal Bacteroides C. jejuni Cer
fragilis expresses polysaccharide a, which, in animal models
of colitis, has been shown to prevent intestinal Neuron cell
inflammation155. Moreover, the glycosylation of the human
intestinal mucus can be altered by the microorganisms that Gal Glc GalNAc Neu5Ac
are present in the gut, for example, through modification
R = any underlying linkage
of α(1,2)fucosylation156–158.
Cer, ceramide; Gal, galactose; Glc, glucose, GalNac, N-​acetylgalactosamine; Neu5ac, N-​acetylneuraminic acid.
in the N-​glycosylation pathway have many phenotypic as carbon sources16,17 to altering host glycans to enable
changes, including reduced mouse75 and chicken colo- cell adhesion81. The most widely studied group of these
nization76, reduced adherence and invasion of gut epi- modifying enzymes are the neuraminidases — enzymes
thelial cells in vitro75, and altered immune-​reactivity72. that cleave and release sialic acid from glycans. Many
Gut proteases can cleave bacterial proteins and attenuate different bacteria produce neuraminidases with varying
the growth of C. jejuni77. Further studies demonstrated specificities82,83. Streptococcus pneumoniae produces over
that N-​linked protein glycosylation protects bacte- ten different glycosidases, including three different neu-
rial proteins from cleavage by gut proteases (Fig. 2d), raminidases with crucial roles in nutrient acquisition and
demonstrating a further strategy of immune avoidance. pathogenesis16. The neuraminidases N-​acetylneuraminate
Nontypeable Haemophilus influenzae also uses glycosylation lyase (NanA), sialidase B (NanB) and neuramidase C
to protect the surface-​exposed high molecular weight (NanC) are important for S. pneumoniae colonization
adhesin 1 (HMW1A). The HMW1C glycosyltransferase as they provide a carbon source from the host mucosa
glycosylates HMW1A, which protects HMW1A from and have a central role in biofilm formation16,84,85.
degradation and anchors the protein to the bacterial S. pneumoniae is a common cause of secondary infection
surface78,79. Recently, O-​linked glycosylation of flagella of in cases of influenza A virus infection86. The action of influ-
several species has been identified as crucial for damp- enza A neuraminidase on host cell sialylated structures
ening immune responses. For example, in Burkholderia promotes subsequent or co-​infection by S. pneumoniae86–88.
cenocepacia the addition of a single 4,6-dideoxy-4- Although bacterial neuraminidases have been
(3-hydroxybutanoylamino)-d​-glucose in multiple places widely studied, other bacterial glycosidases, such as
on the flagellum reduces Toll-​like receptor 5 signalling80. heparanases, have not been studied to the same extent.
For example, the lack of structural information on
Host protein modifications. Bacterial pathogens modify heparanases has hindered structure–function studies.
host glycoconjugates using a range of glycosyltransferases However, a recent study determined the structure of the
and glycosidases. The modification of host glycocon- Burkholderia pseudomallei heparanase89, which may pro-
jugates provides the bacterium with a range of host vide insights into substrate recognition that could enable
adaptation functions, ranging from accessing glycans future studies of novel heparanase inhibitors89.
© 2018 Macmillan Publishers Limited, part of Springer Nature. All rights reserved.
Reviews

a Macrophage

β3 β4 β3 β4
GAS LMW HA

β3 β4 β3 β4 β3 β4 β3 β4 Epithelial cell
GAS HMW HA

CD44

b
Siglec-9
β4 α3
GBS

Neutrophil

β3 β4 β3 β4
GAS

c
C3b Alternative Alternative
complement complement

N. meningitidis IS1301

d
Gut protease α3 α4 α4 α4 α4
N
β3

C. jejuni

e EPEC Host
Death domain receptor

FADD Caspase 8
Type III
secretion NleB1
system
Apoptosis

Gal Glc GalNAc GlcNAc Neu5Ac GlcA Bac

Fig. 2 | Glycointeractomics of pathogen persistence. a | The molecular weight of the hyaluronan (HA) expressed on the
group A Streptococcus (GAS) polysaccharide capsule has roles in immune evasion. Low molecular weight (LMW) HA leads to
greater uptake by macrophages, whereas the high molecular weight (HMW) HA capsule interacts with CD44 on host cells
and has less uptake by macrophages. b | Both group B Streptococcus (GBS) and GAS have polysaccharide capsules, with the
GBS capsule comprising Siaα2-3Galβ1-4GlcNAc and the GAS capsule comprising HA. Both capsular polysaccharides
interact with sialic acid-​binding immunoglobulin-​like lectin 9 (Siglec-9) as a method of immune evasion but at two different
binding sites. The site of interaction between GBS Siaα2-3Galβ1-4GlcNAc capsule and Siglec-9 is known, but the site of HA
on Siglec-9 is unknown. c | An insertion sequence (IS1301) between the sia and ctr operons, which are responsible for the
biosynthesis and export of the capsule, causes higher levels of capsular polysaccharide to be expressed in Neisseria
meningitidis serogroup C. The increased capsular polysaccharide expression leads to impairment of the alternative
complement pathway and vaccine escape. d | Host proteases can act as a part of the non-​adaptive immune system by
destroying surface proteins of pathogenic bacteria. The N-​linked protein glycosylation of Campylobacter jejuni protects
bacterial proteins from cleavage by host gut proteases. e | The type III secretion system of enteropathogenic Escherichia coli
(EPEC) delivers the effector protein NleB1, which acts as an N-​acetylglucosamine (GlcNAc) transferase to modify FAS-​
associated death domain protein (FADD). This glycosylation of a host protein by EPEC avoids caspase 8 activation and as
such prevents host cell death receptor apoptosis. Bac, bacillosamine; Gal, galctose; GalNAc, N-​acetylgalactosamine; Glc,
glucose; GlcA , glucuronic acid; Neu5Ac, N-​acetylneuraminic acid. Sia, sialic acid.

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Type III secretion system
A needle-​like structure
produced by pathogenic
bacteria to deliver proteins to
the host cell to facilitate
bacterial infection.
Death receptor-​mediated
apoptosis
Also known as extrinsic
apoptosis; programmed cell
death activated by immune
system signals.
AB5 class toxins
Toxins comprising one A
subunit that corrupts critical
cell function and five B
subunits that direct target
cell uptake.
B subunits
Subunits of an AB toxin that
are responsible for binding
to host cells.
A subunits
Subunits of an AB toxin that
are enzymatically active.
Surface plasmon resonance
An optical technique for the
measurement of adsorption
of materials onto a surface
that can estimate affinities
of proteins for glycans on
coated chips.
Glycomics
Systematic analysis of the
glycome.
Isothermal titration
calorimetry
A technique that measures the
heat released or absorbed
during the binding of molecules
to determine the binding
constants, reaction
stoichiometry, enthalpy and
entropy of the interaction.
Saturation transfer
difference NMR
A NMR technique for
determining the interaction
between proteins and small
molecules by the transfer of
magnetic field from the protein
to the small molecule.
Nature Reviews | MicrobioloGy
A novel mechanism for glycosylation of a host
protein was recently reported. Enteropathogenic
E. coli (EPEC) has a type III secretion system that is used
to deliver effector proteins into the host cell, includ-
ing NleB1, a N-​acetylglucosamine transferase90,91. The
effector protein NleB1 modifies the adaptor protein
FAS-​associated death domain protein (FADD), which
is involved in host cell apoptosis after infection, by addi-
tion of a GlcNAc residue at a specific arginine92 (Fig. 2e).
Glycosylation at this location of FADD prevents death
receptor signalling and the activation of caspase 8,
which is required for death receptor-​mediated-​apoptosis92.
The GlcNAcylation of the arginine is a non-​reversible
glycosylation event, as mammalian enzymes are unable
to remove this carbohydrate residue92. Glycosylation to
prevent apoptosis represents a novel virulence and
immune-​avoidance strategy.
Toxins
Host glycans are common targets for many bacte-
rial toxins. Some of the best-​studied toxins are the
AB5 class toxins from Vibrio cholerae (cholera toxin),
Bordetella pertussis (pertussis toxin) and Shigella dys-
enteriae and Shiga toxigenic E. coli (STEC) (Shiga toxin)
and neurotoxins from Clostridium botulinum (botu-
linum toxin) and Clostridium tetani (tetanus toxin).
The B subunits of these toxins mediate binding to tar-
get glycans on cell surfaces and direct the uptake and
intracellular trafficking of the toxin93,94. Many of these
toxin–glycan interactions have been well characterized
and have been reviewed elsewhere93–96. However, several
new toxin–glycan interactions were recently identified
and are discussed below.
AB5 toxin SubAB. Apart from Shiga toxin, some STEC
strains express an additional AB5 toxin known as the
subtilase cytotoxin (SubAB)97. The SubAB binding sub-
unit SubB binds glycans terminating in the sialic acid N-​
glycolylneuraminic acid (Neu5Gc) and exhibits a strong
binding preference for Neu5Gc over Neu5Ac18 (Fig. 3a),
thus the tropism for this toxin are cells that express
Neu5Gc. Unlike most mammals98,99, humans do not pro-
duce Neu5Gc, as they lack the cytodine monophosphate-​
N-acetylneuraminic acid hydroxylase (CMAH) protein
— the enzyme that converts Neu5Ac to Neu5Gc100.
Instead, humans acquire Neu5Gc from their diet101 and
incorporate it into gut epithelia and vascular endothelia,
and in doing so, sensitize these tissues to this toxin18,102.
CdtB, PltA and PltB. Salmonella enterica subsp. enterica
serovar Typhi (S. Typhi), unlike other Salmonella enterica
serovars, is a human-​exclusive pathogen19,103,104. S. Typhi
is the cause of typhoid fever, and the typhoid toxin is the
cause of most of the symptoms of the disease. This toxin
is a member of the AB5 toxin family, but it is unique in
that there are two A subunits, cytolethal distending toxin
subunit B homologue (CdtB) and pertussis-​like toxin
subunit A (PltA); thus this toxin defines a new subclass
of AB5 toxin, A2B5 (ref.105). Cytotoxic assays with either
Neu5Ac-​expressing or Neu5Gc-​expressing cells showed
that typhoid toxin was only cytotoxic to cells expressing
Neu5Ac. Mice that were engineered to constitutively
© 2018 Macmillan Publishers Limited, part of Springer Nature. All rights reserved.
Reviews
express CMAH, therefore presenting Neu5Gc, are resistant
to typhoid toxin. This study demonstrated that the toxin
has a preference for the sialic acid Neu5Ac over Neu5Gc19
(Fig. 3b). This preference is thought to contribute to the
human-​specific nature of S. Typhi infections19.
Pneumolysin: a cholesterol-​dependent cytolysin. S.
pneumoniae is a major cause of pneumonia, septicaemia
and meningitis in humans20. The vast majority of clin-
ical isolates of S. pneumoniae produce the cholesterol-​
dependent cytolysin (CDC), pneumolysin (Ply)106. CDCs
are pore-​forming toxins that require membrane choles-
terol for cytolytic activity20. Given the ubiquitous presence
of cholesterol in mammalian cell membranes, binding of
cholesterol cannot fully explain the cell tropism of Ply for
red blood cells and phagocytic white cells20. Formation of
the trans-​membrane pore is a multistep process: attach-
ment of domain 4 of the toxin to the host cell, oligomeri-
zation and penetration of the host cell membrane, leading
to the formation of a trans-​membrane β-​barrel pore107.
Glycan microarray analysis (see Glycan microarray
section) was used to identify sLeX and Lewis X as cel-
lular receptors for Ply (Fig. 3c). Surface plasmon resonance
was used to confirm the microarray results, and it also
showed that interactions with both glycans were of high
affinity20. The haemolytic activity of Ply can be inhibited
by sLeX. Moreover, free sLeX blocks attachment of the
toxin to the red blood cell surface20. In silico analyses
suggested that glycan binding involved domain 4 of Ply,
which was confirmed by mutagenesis of the putative
glycan-​binding amino acids20. These data indicate that
in addition to cholesterol, Ply has a glycan cell receptor
that contributes to target cell tropism.
Glycomics and glycointeractomics
The increase in the number of publications on glycoint-
eractomics in the past five years is in part due to technical
advances, in particular the application of microarray plat-
form technologies to glycomics arrays4–6,108–114. The availa-
bility of high-​throughput screening platforms has led to a
major increase in the understanding of glycointeractomes.
Microarray technology is based on the theory that a small
high-​density spot of a bioactive molecule provides greater
sensitivity compared with conventional immunoassay
formats, as miniaturized features can increase detection
sensitivity115. A microarray is composed of a screening
panel of compounds that are printed onto a solid support
matrix, often a functionalized glass slide109. Glycan, lectin
and mucin microarray technologies are discussed in detail
below. These technologies are currently the only high-​
throughput methods that are available for the analysis
of glycointeractions. Once discovered, glycointeractions
require validation and quantification using complemen-
tary methods, such as isothermal titration calorimetry and
surface plasmon resonance116,117, and by characterization of
the interactions at the atomic level using techniques such
as NMR approaches. Saturation transfer difference NMR in
particular has emerged as a powerful tool to study gly-
cointeractions. In a recent study, interactions between
influenza A virus receptor glycans and whole viral par-
ticles were reported98,118. The optimal technical approach
for the detection of glycointeractions would be a system in
Reviews
Anomeric configurations
The bond angle on carbon-1
for most saccharides or
carbon-2 for sialic acids (that
is, α or β linkage).
a b c
SubAB PltB Ply
(from Cdtb–PltA–PltB)
sLeX
Neu5Ac Neu5Gc Neu5Ac Neu5Gc
α3
α3 α3 α3 α3 β4
β4 β4 β4 α3 β4 α3 α3
β3
β3 β3 Toxin pore
R R R R R
R R R R
Red blood cell
Gut epithelial cell
Gal Glc GlcNAc Fuc Neu5Ac Neu5Gc R = any underlying linkage
Fig. 3 | Glycointeractomics of bacterial toxins. a | The B subunits (boxed in red) of the AB5 toxin subtilase cytotoxin
(SubAB, PDB ID 4BWG165) strongly bind glycans terminating in N-​glycolylneuraminic acid (Neu5Gc) and intoxicate host
cells expressing Neu5Gc over N-​acetylneuraminic acid (Neu5Ac). b | Pertussis-​like toxin subunit B PltB (PDB ID 4RHR19), the
B subunit of typhoid toxin cytolethal distending toxin subunit B homologue (CdtB)–PltA–PltB, has a preference for Neu5Ac,
the sialic acid expressed by humans, over Neu5Gc. c | Domain 4 of the cholesterol-​dependent cytolysin pneumolysin
(Ply ; PDB ID 4QQA) (boxed in red) binds to sialyl Lewis X (sLeX) on the surface of red blood cells. This attachment allows the
oligomerization of the toxin to form a pore in the red blood cell membrane. Fuc, fucose; Gal, galctose; Glc, glucose; GlcNAc,
N-​acetylglucosamine; Neu5Ac, N-​acetylneuraminic acid; Neu5Gc, N-​glycolylneuraminic acid.
which the glycan microarray platform can simultaneously density of a specific lectin in spots on the microarray
detect binding and determine the kinetics of the inter- matrix provides multivalency effects that more closely
action. Efforts to achieve this goal are in development reflect the native presentations. Multivalency is a core
using several platforms, including the combined quartz property of lectins that is used to increase specificity and
crystal microbalance and microarray that were recently avidity of binding. The binding specificities of a number
described119. of lectins have been characterized121. This information
It is important to emphasize the distinction between makes it possible to determine the monosaccharide types,
approaches that detect glycointeractions that underpin linkages and anomeric configurations of a glycan and thus
the identity of structures in the sample being analysed.
glycointeractomics and techniques that separate, identify
Currently, the structural information is mostly limited to
and/or quantify glycans in the glycomics analysis of bio-
logical samples120 (Box 3). Glycomics approaches, such as
terminal structures122. Current lectin microarrays con-
high-​performance liquid chromatography (HPLC) and tain up to 96 glycan-​binding proteins3–5,113. Limitations
mass spectrometry, are destructive analyses, requiring of lectin microarray technology, including glycan struc-
the release of the glycans from the biological source. tural elucidation, are limited to precise definition of the
Moreover, these techniques are not routinely used to structural epitopes that are recognized by lectins and the
measure interactions between cell surface glycans and limited availability of commercially available lectins to
expand the structural repertoire that can be detected114.
bacterial lectins or vice versa. Therefore, these techniques
currently have limited use in glycointeractomics studies. Lectin microarrays have been used to analyse a
range of biological systems from human tissue to
Lectin microarrays. Lectins are proteins that preferen- bacteria (Fig. 4a). The major application has been to
tially recognize and bind carbohydrates. These glycan-​ investigate the glycosylation of purified proteins or
binding proteins not only bind glycans on cell surfaces whole-​cell glycosylation states. Lectin microarrays
but also bind free carbohydrates120. Some lectin–glycan have been used to evaluate the surface glycosylation
interactions can be low-​affinity, and printing a high of pathogenic and non-​pathogenic E. coli strains. The
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pathogenic E. coli neonatal meningitis strain RS218
had a distinct binding pattern from the non-​pathogenic
K12 strains JM101 and HB101 and expressed a wider
range of surface glycan structures123. LOS expressed
on the surface of C. jejuni has been analysed by lectin
microarray, and it confirmed the predictive power of
the genetic composition of the LOS biosynthesis clus-
ters3. Lectin microarrays have also been used to deter-
mine the glycosylation patterns of 24 commonly used
mammalian cell lines4.
Glycan microarrays. Glycan microarrays are glycan-​
binding assays implemented in a sensitive format that
utilizes a small amount of sample and permits the
simultaneous analysis of binding to hundreds of test
glycans in one high-​throughput assay124–126 (Fig. 4b). The
utility of the glycan microarray is linked to the num-
ber of structurally defined glycans that are present on
the array. As such, the most useful glycan microarray
would contain all glycans in a glycome; however, most
glycan microarrays currently have ~500 glycans that are
derived mostly from mammals7,127,128 compared with
the many thousands of possible glycans in nature129.
Pathogens usually bind carbohydrates on the host cell
surface, and the glycan microarray provides a screen-
ing tool to discover the glycan-​binding capabilities of
various microorganisms124. A C. jejuni mutant lacking
the chemotaxis receptor tlp11 showed reduced virulence
in a chicken colonization model130. Glycan microarrays
showed that C. jejuni Tlp11 specifically interacts with
galactose, regardless of whether the galactose was a free
monosaccharide or the terminal sugar of a larger oligo-
saccharide or polysaccharide130. Identification of carbo-
hydrate ligands for a range of proteins, toxins and whole
bacteria using glycan microarray screening was also per-
formed for streptococcal M protein11 SubB18, typhoid
toxins cytolethal distending toxin subunit B homolog
CdtB, PltA and PltB19, and Ply20. Glycan microarrays
have also been used to discover novel glycan–glycan
interactions between bacteria and host cells116,117 (Box 1).
Recently, a novel glycan microarray was developed
with defined glycans from capsular polysaccharides,
LPS and LOS from a range of bacteria6. The bacterial
polysaccharides were immobilized onto a functionalized
glass slide. This microarray was used to test the immune
response to a challenge from a range of bacterial spe-
cies. These studies showed that the immunoglobulins
produced to each pathogen had unique patterns of reac-
tivity to glycans depending on the infected mammalian
species6. This microarray was also used to show that
host galectins have specificity for microbial glycans that
mimic host antigens. For example, galectin 3, galectin
4 and galectin 8 bound the O-antigen of Providencia
alcalifaciens, which contains the mammalian α-​galactose
epitope (Galα1-3 Gal)6.
Mucin microarrays. Epithelial cells within the body are
coated with mucus, and mucin is the major component
of these mucous layers131. Mucins are high molecular
weight glycoproteins that can be membrane-​bound or
secreted and are heavily glycosylated with 50–90% carbo-
hydrate by mass109. The glycan structures on mucins are
Nature Reviews | MicrobioloGy
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Reviews
Box 3 | Defining a glycome
One of the most crucial steps for understanding the
glycointeractome is to be able to accurately define the
glycome of both the host and the pathogen. the two
most commonly used methods to define the glycome are
high-​performance liquid chromatography (HPLC)120,159
and mass spectrometry120,160. HPLC is a separation
technique that can be used for both neutral and charged
glycans. separation by HPLC can be performed in various
ways, including different anion exchange methods and
the use of normal or reverse phase separation120,159. there
are disadvantages of HPLC; for example, determining the
sequence of monosaccharides in an oligosaccharide or
polysaccharide is difficult owing to similarities between
glycans, and complex samples often provide data peaks
that may contain more than one glycan. HPLC does not
yield detailed structural information, but it can be used
in conjunction with mass spectrometry to gain structural
information. Mass spectrometry is a good choice for
analysing very complex glycan mixtures containing
unusual carbohydrate structures160. it operates by
ionizing compounds to create charged molecules and
separates the molecules by mass-​to-charge ratio,
displaying the separated charged molecules that are
detected as a spectrum. the molecules are identified by
correlating known masses or characteristic
fragmentation patterns of the spectral data160–162. recent
advances in mass spectrometry have included the
analysis of glycopeptides, giving the location of precise
glycosylation sites on a glycoprotein163, and imaging
mass spectrometry, which can provide information on
the cellular and tissue distribution of glycans164. Neither
of these techniques have been applied to bacterial
pathogens, but they may become powerful tools for
understanding host–pathogen interactions in the future.
highly heterogeneous. They can vary on the same mucin
and vary between tissue locations and during health and
disease states131,132. Yields of purified mucins from biolog-
ical samples are often low, making the high-​throughput
method of microarrays ideal for studying interactions
between mucins and bacteria. An array of 35 naturally
sourced purified mucins was constructed124, with the
mucins sourced from the mucosal layers of the gastro-
intestinal, respiratory and reproductive tract of different
animal species and from human gastrointestinal tract cell
lines. The purified mucins were conjugated to a function-
alized hydrogel glass slide surface and then probed with
lectins to elucidate the glycan structures present on the
mucins. Distinct mucin glycan structures were reported
from mucins isolated from different animals and tis-
sues109,132. In a separate study, the mucin array was used to
study interactions between mucins and whole bacteria8.
A range of H. pylori and C. jejuni strains were found to
bind different mucins, with the C. jejuni strains binding
strongly to mucins from chicken intestines and caecum,
whereas H. pylori bound to several mucins, including
those from porcine stomach and chickens8.
Computational methods. Another glycointeractomics
approach utilizes computational methods to identify
glycan-​binding proteins. As previously discussed, bac-
terial glycan-​binding adhesins are important for adher-
ence, colonization and persistence. As such, bacterial
Reviews

similarity for those proteins that are not necessary

similar in sequences. Another recent method called
a Lectin array sequence-​based prediction of residue-​level interactions
sites of carbohydrates (SPRINT-​CBH) trained on known
lectin–carbohydrate interactions by machine-​learning
for the computational identification of novel lectins136
using sequence-​derived information alone. SPRINT-​CBH
Printed
lectins uses over 1,000 protein–carbohydrate complex structures
compiled from a range of databases, including Glyco3d
and PROCARB, to provide training sequences for lectin
site predictions136. This method is not limited to proteins
with known structures and thus can be used in a genome-​
Cell Toxin Bacteria
Printed scale prediction. In summary, computational methods
glycans are being developed to identify glycan-​binding proteins
using protein sequence and structure (if known). These
methods employed currently known complex databases
and are expected to become more accurate if additional
b Glycan array information generated from other glycomic and glycoin-
teractomic techniques can be added to the databases for
training computational methods.

Fig. 4 | Applications of lectin and glycan microarrays in glycointeractomics. Conclusions

a | Lectins are carbohydrate-​binding proteins with known binding specificities for glycan Glycointeractions between the host and pathogenic
structures. Current lectin microarrays have up to 96 lectins printed in replicate, providing bacteria have a crucial role during all stages of infec-
coverage of most glycans known to be expressed on humans and pathogenic bacteria. tion. During bacterial colonization, pathogenic bacterial
These microarrays have been used to discover what glycan structures are available on the adhesins can act as lectins, as observed with the adhesins
surface of cells, bacteria and bacterial glycoproteins. b | Glycan (and mucin) microarrays in E. coli and H. pylori, or, conversely, bacteria may use a
are used to define the glycan-​binding abilities of cells, bacteria and proteins, including host lectin for adherence, as reported in N. gonorrhoeae
toxins. Discovering which glycans are available to bind is an important step in defining
infection. To evade the immune system and persist
the glycointeractome between the host and pathogen.
within the host, pathogenic bacteria can express glycans
that mimic host structures, as shown with GAS and HA.
lectins are potential therapeutic targets133. In silico Glycointeractions have a central role in pathology, with
analyses can be used to screen bacterial proteins of many bacterial toxins displaying cell, tissue and species
unknown function for potential carbohydrate-​binding tropism based on the presentation of host cell glycans
properties, thereby providing another method for as cellular receptors. Elucidating the glycointeractome
identifying novel bacterial lectins. One method of in is an essential step in understanding how pathogenic
silico analysis involves a structure-​based function pre- bacteria interact with the host, and approaches to fully
diction programme called SPOT-​Struc, where glycan-​ define the glycointeractome have developed rapidly in
binding proteins are identified by structural similarity recent years. An important area that requires further
to known lectins (templates) in a database134. This pro- effort is the development of comprehensive glycobio-
gramme identifies carbohydrate-​binding proteins and informatics resources that can be used to exploit gly-
recognition sites by using the structural alignment comic data by linking glycan structure, expression and
tool SPalign134,135. Well-​a ligned structures are then function. The developing field of glycointeractomics
used to model complex structures and predict bind- has great potential to resolve fundamental aspects of
ing affinities between a query protein structure and bacterial pathogenesis and present opportunities for
the template glycan. Calculated binding affinities are novel therapeutic and preventive strategies to combat
then used to filter results and remove potential false bacterial infections.
positives of carbohydrate-​binding proteins134. Such a
Published online xx xx xxxx
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Acknowledgements
This work was supported by a National Health and Medical
Research Council (NHMRC) Program Grant 1071659 to J.C.P.,
M.v.I. and M.P.J., Principal Research Fellowship 1138466 to
M.P.J. and Project Grant 1108124 to M.P.J. and C.J.D.
Reviews
Author contributions
J.P., C.J.D., M.v.I., J.C.P. and M.P.J. conducted the literature
review for this article. J.P., C.J.D., M.v.I., J.C.P. and M.P.J.
substantially contributed to discussion of content and wrote
the article. J.P., C.J.D. and M.P.J. designed the figures. J.P.,
C.J.D., J.C.P. and M.P.J. reviewed and edited the manuscript
before submission.
Competing interests
The authors declare no competing interests.
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