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Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology. 2nd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2009.

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Essentials of Glycobiology. 2nd edition.

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Chapter 13Structures Common to Different Glycans

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In the section “Poly-N-acetyllactosamines,” the following sentence was deleted: “Glycoproteins and glycolipids frequently bear N-glycans, O-glycans, and glycolipids with poly-N-acetyllactosamine chains.”

This chapter describes terminal glycan structures that are common to different classes of glycans. They are the variable portions of N-glycans, O-glycans, and glycolipids that are attached to the core sugars characteristic of each glycan class. The core structures of N-glycans, O-glycans, and glycolipids (Figure 13.1) are the product of biosynthetic pathways discussed in Chapters 8, 9, and 10. More complicated cores can result from tissue- or cell-type-specific pathways and lead to further structural diversification of glycan chains. The glycan units formed by subultimate and terminal sugars at “outer” positions of a glycan often determine the function(s) or recognition properties of a glycoconjugate.

FIGURE 13.1. N-Glycan synthesis leads to structures (a biantennary N-glycan in this example; see Chapter 8) that are subsequently modified by branching N-acetylglucosamine (GlcNAc) residues and other sugars (arrows) in glycosylation reactions that may be tissue-specific, developmentally regulated, or even protein-specific.


N-Glycan synthesis leads to structures (a biantennary N-glycan in this example; see Chapter 8) that are subsequently modified by branching N-acetylglucosamine (GlcNAc) residues and other sugars (arrows) in glycosylation reactions that may be tissue-specific, (more...)


In contrast to core glycan synthesis, which is constitutive in most cell types, the addition of branching and terminal sugars is often regulated in a tissue- or cell lineage–specific manner (Figure 13.1). Many of these reactions are regulated during embryogenesis and in the postnatal period as part of the normal developmental program (see Chapter 38). Changes in terminal glycan structure are also often associated with malignant transformation in cancer (see Chapter 44). As discussed in Chapters 5 and 38, tissue- and/or lineage-specific regulation of outer-chain biosynthesis is largely a function of the regulated expression of the relevant glycosyltransferases. Biological consequences that correlate with a number of changes in outer-glycan units are discussed throughout this volume. However, the majority of regulated terminal glycosylations observed do not yet have a defined function.

Type-2 Glycan Units

The more mature glycans shown in Figure 13.1 contain terminal N-acetylglucosamine (GlcNAc) residues. The subsequent addition of galactose in β1–4 linkage generates a type-2 unit, which is composed of the disaccharide Galβ1–4GlcNAc, also called N-acetyllactosamine (LacNAc) (Figure 13.2). The terminal galactose so generated can be modified by the addition of a GlcNAc residue, which in turn can receive a galactose in β1–4 linkage, thus forming two LacNAc units. These reactions may continue to form poly-N-acetyllactosamine [Galβ 1–4GlcNAc]n. Poly-N-acetyllactosamine chains are found in glycans from most cell types. An alternative to type-2 LacNAc units is a chain composed of so-called LacdiNAc glycan units generated by the action of a β1–4 N-acetylgalactosaminyltransferase, to form [GalNAcβ 1–4GlcNAc]n. Efforts are being made to define the biological role(s) for type-2 chains by the generation and analysis of mice deficient in β1–4 galactosyltransferase genes, of which there are six that can generate type-2 chains.

FIGURE 13.2. Terminal GlcNAc residues are usually galactosylated.


Terminal GlcNAc residues are usually galactosylated. Modification by β1–4Gal residues (top) occurs in all mammalian tissues. This reaction is catalyzed by β1–4 galactosyltransferase (β1–4GalT) and yields (more...)

Type-1 Glycan Units

GlcNAc residues in N-glycans, O-glycans, and glycolipids may instead be modified by galactose in β1–3 linkage (Figure 13.2) to obtain a type-1 unit composed of a disaccharide termed neo-N-acetyllactosamine. In humans, expression of type-1 units is mostly restricted to epithelia of the gastrointestinal or reproductive tracts. Type-1 units may be modified by glycosyltransferases that transfer sugars to either terminal galactose or subterminal GlcNAc, generating sialylated structures or blood group determinants. The synthesis of type-1 units and expression of the corresponding β1–3 galactosyltransferases are regulated in a tissue-specific manner, but biological roles for this disaccharide remain to be defined.


As noted above, poly-N-acetyllactosamine biosynthesis is directed by the alternating actions of β1–4 galactosyltransferases and β1–3 N-acetylglucosaminyltransferases (Figure 13.2). Some glycoproteins and glycolipids are preferentially modified to carry poly-N-acetyllactosamine. This implies that the glycosyltransferases responsible for poly-N-acetyllactosamine biosynthesis can discriminate between glycoproteins or glycolipids that appear to present the same terminal GlcNAc and galactose glycan acceptors. For example, poly-N-acetyllactosamine extensions preferentially occur on multi-antennary N-glycans and particularly on the β1–6GlcNAc branch, the synthesis of which is under the control of GlcNAc transferase V (GlcNAcT-V) (Figure 13.3; see Chapter 8). Similarly, poly-N-acetyllactosamine extensions on O-glycans associated with mucin glycoproteins often preferentially occur on the β1–6GlcNAc transferred by a core 2 β1–6GlcNAcT (Figure 13.3). Poly-N-acetyllactosamine chain length is also under regulatory control, with N-glycans generally having longer chains than O-glycans. Poly-N-acetyllactosamine chains serve as acceptors for subsequent glycosylations, including fucosylation and sialylation. The linear nature of these chains together with their hydrophilic character predicts that they have an extended linear conformation. Thus, poly-N-acetyllactosamine chains may serve as scaffolds for the presentation of specific terminal glycans, whose functions require them to be presented at a certain distance from the plasma membrane. Poly-N-acetyllactosamine chains are also recognized with high affinity by mammalian galectins (see Chapter 33) in a manner that implies recognition of internal sugars as well as terminal galactose.

FIGURE 13.3. Poly-N-acetyllactosamine chains are found on N-glycans, O-glycans, and glycolipids.


Poly-N-acetyllactosamine chains are found on N-glycans, O-glycans, and glycolipids. LN designates N-acetyllactosamine unit. Symbol Key: Image symbol_key_small.jpg

β 1-6GlcNAc-branched Poly-N-acetyllactosamines

Poly-N-acetyllactosamine chains may be branched by the addition of GlcNAc in β1-6 linkage to internal galactose residues through the action of β1-6 N-acetylglucosaminyltransferases (β1–6GlcNAcTs) (Figure 13.4). Branched and nonbranched poly-N-acetyllactosamine chains correspond to the “I” and “i” antigens, respectively. These antigens were discovered by the analysis of a cold-dependent agglutinating antibody (cold agglutinin) in a patient with acquired hemolytic anemia (see Chapter 43). Cold agglutinin antibodies react with red blood cells of most blood donors. Nonreactive donors are classified as having the i blood group, whereas reactive donors are assigned to the I blood group. Thus, β1–6-branched poly-N-acetyllactosamine units correspond to the I blood group antigen and linear poly-N-acetyllactosamine chains that lack branching correspond to the i blood group antigen. Distinct β1–6GlcNAcTs yield different types of β1–6-branched structures (Figure 13.4). Glycans reactive to I and i blood group antibodies are expressed by many human cells. The i antigen is abundantly expressed on the surface of embryonic red cells and on red cells during times of altered erythropoiesis. Such cells are relatively deficient in the expression of the I antigen. However, during the first 18 months of life, I antigen reactivity on red cells reaches adult levels, and i antigen reactivity declines to very low levels. This developmental regulation is presumed to be due to regulated expression of I β1–6GlcNAcT genes. Rare individuals never express the I antigen on red cells and maintain embryonic levels of red cell i antigen expression as adults. Individuals with this i phenotype may be homozygous for inactive alleles at β1–6GlcNAcT loci. There is no obvious pathophysiology associated as yet with the absence of the I blood group.

FIGURE 13.4. i and I antigen synthesis.


i and I antigen synthesis. Linear poly-N-acetyllactosamine chains (i antigen) synthesized on N- and O-glycans or glycolipids (R) may be modified by β1–6 N-acetylglucosaminyltransferases (β1–6GlcNAcTs). These enzymes transfer (more...)


In humans, poly-N-acetyllactosamine chains and their β1–6-branched variants are subject to tissue-specific glycosylations that form the ABO blood group antigens. The ABO system was discovered early in the twentieth century by Karl Landsteiner and colleagues. They showed that humans could be divided into different classes according to the presence or absence of serum factors that would agglutinate red cells isolated from other humans. We now know that these serum factors are antibodies and that the corresponding antigens are glycan epitopes, whose structures are determined by the inheritance of genes that encode glycosyltransferases with different activities.

The A, B, and H blood group antigens are glycans presented on the type-1 or type-2 N-acetyllactosamines described above (Figure 13.5), on O-GalNAc glycans (type-3; Figure 13.6), or on glycolipids (type-4; Figure 13.7). The A, B, and H antigens are formed by the sequential action of glycosyltransferases encoded by three genetic loci (the ABO, H, and Secretor [Se] loci) (Figure 13.8). The pathway of ABO blood group antigen synthesis begins with modification of type-1 or type-2 N-acetyllactosamine by α1–2 fucosyltransferases (α1–2FucTs). The transfer of fucose in α1–2 linkage to the galactose in type-1 or type-2 N-acetyllactosamine forms the blood group H determinant. The human genome encodes two different α1–2FucTs, encoded by the H and Se blood group loci, respectively. The H α1–2FucT is expressed in red cell precursors, and it transfers fucose to type-2 and type-4 glycan units to form the H antigen on red cells (Figure 13.5 and Figure 13.7). The Se α1–2FucT is expressed in epithelial cells, and it uses type-1 and type-3 N-acetyllactosamines to form the H antigen (Figure 13.5 and Figure 13.6) in epithelia lining the lumen of the gastrointestinal, respiratory, and reproductive tracts and in salivary glands.

FIGURE 13.5. Type-1 and -2 H, A, and B antigens that form the O (H), A, and B blood group determinants on N- and O-glycans.


Type-1 and -2 H, A, and B antigens that form the O (H), A, and B blood group determinants on N- and O-glycans. Symbol Key: Image symbol_key_small.jpg

FIGURE 13.6. Type-3 H, A, and B antigens that form the O (H), A, and B blood group determinants on N- and O-glycans.


Type-3 H, A, and B antigens that form the O (H), A, and B blood group determinants on N- and O-glycans. Symbol Key: Image symbol_key_small.jpg

FIGURE 13.7. Type-4 H, A, and B antigens that form the O (H), A, and B blood group determinants on glycolipids.


Type-4 H, A, and B antigens that form the O (H), A, and B blood group determinants on glycolipids. Symbol Key: Image symbol_key_small.jpg

FIGURE 13.8. Synthesis of H (O), A, and B blood group determinants.


Synthesis of H (O), A, and B blood group determinants. For details, see text. Symbol Key: Image symbol_key_small.jpg

A or B blood group determinants are subsequently formed from type-1, -2, -3, or -4 H determinants by glycosyltransferases encoded by the ABO blood group locus. The blood group A glycan epitope is formed by the α1–3GalNAcT encoded by the A allele of the ABO locus (Figure 13.8). The blood group B allele of the ABO locus encodes the α1–3GalT that forms the blood group B glycan determinant (Figure 13.8). O alleles at the ABO locus encode a functionally inactive A/B glycosyltransferase. Individuals who synthesize exclusively A determinants are blood group A and have the genotype AA or AO, whereas blood group B individuals have the genotype BB or BO. Individuals who express both A and B determinants are blood group AB and have the genotype AB. Blood group O individuals do not express either A or B determinants (their H antigen is unmodified). They are homozygous for the inactive O allele and their genotype is OO.

The ABO antigens are expressed on membrane glycoproteins and glycolipids on the surface of red cells and other cells in many tissues, including the vascular endothelium and a variety of epithelia. Some tissues also synthesize soluble, secreted forms of these molecules as glycans on secreted glycoproteins, glycolipids, and free glycans. As discussed below, the ability to secrete soluble molecules carrying ABH blood group antigens is a genetically determined trait that is a function of alleles at the Se locus.

On each human red blood cell, approximately 1–2 million ABH determinants (~80% of the total) are attached to the anion transport protein (also known as band 3). About half a million ABH determinants are carried by the red cell glucose transport protein (band 4.5). Both of these integral membrane proteins carry ABH antigens on a single branched N-glycan with poly-N-acetyllactosamines whose terminal branches may carry several ABH determinants. Small numbers of ABH antigens are expressed by other red cell glycoproteins. Each red cell also expresses approximately half a million glycolipids with ABH determinants. Many of these glycolipids have A, B, and H determinants on poly-N-acetyllactosamine chains and have been termed polyglycosylceramides or macroglycolipids. A, B, and H determinants based on type-4 chains (Figure 13.7) are also present in human red cell glycolipids.

ABH determinants expressed by the epidermis are primarily constructed from type-2 units (Figure 13.5). Mucins derived from the gastric mucosa and from ovarian cyst fluid express A, B, and H antigens on type-3 units (Figure 13.6). As noted above, the epithelial cells lining the digestive, respiratory, urinary, and reproductive tracts express type-1 units, as do the epithelia of some salivary and other exocrine glands. These tissues synthesize soluble forms of the ABH determinants, which are therefore largely carried on type-1 units. Expression of the A, B, and H determinants in such secretory tissues is a function of the α1–2FucT encoded by the Se gene, since the H gene is not expressed in these tissues. Humans with an inactive Se gene do not express soluble forms of the A, B, or H determinants in saliva or in other tissues, even though they express the soluble precursors. The term nonsecretor is used to describe these individuals and refers to the fact that soluble blood group A, B, or H antigens cannot be detected in their saliva.

Serology used to characterize red cells for transfusion has identified variants of the A and B blood group determinants that typically yield weak reactivity with typing reagents. For example, the lectin Dolichos biflorus agglutinates red cells from most blood group A individuals (termed A1 individuals), but it does not agglutinate red cells from individuals of the A2 subgroup. The molecular structures of A1 and A2 subgroup antigens are distinct (Figure 13.9). These structural differences reflect the different catalytic activities of the A transferases encoded by the A1 and A2 alleles.

FIGURE 13.9. A1 and A2 blood group antigens.


A1 and A2 blood group antigens. The type-2 A glycan is modified by a β1–3GalT and the α1–2FucT from the H locus to form a type-3 H determinant and the A2 blood group. The type-3 H glycan is a substrate for the A1 transferase (more...)

The heritable red cell antigenic polymorphisms determined by the ABO locus have important medical implications. Early in the postnatal period, the immune system generates IgM antibodies against ABO antigen(s) when they are absent from an individual’s red cells. The antibodies likely reflect an immune response to glycan antigens presented by bacteria and fungi that express glycans similar or identical to the A and B blood group determinants. For instance, type-O individuals do not make A or B determinants and they exhibit relatively high titers of circulating IgM antibodies (termed isoagglutinins) against A or B blood group determinants. Similarly, blood group B individuals exhibit circulating IgM anti-A isoagglutinins, but they do not make isoagglutinins against the blood group B determinant, a “self” antigen. Sera taken from blood group A individuals contain anti-B but not anti-A antibodies. Finally, those individuals with the AB blood group do not make either anti-A or anti-B IgM isoagglutinins. Anti-H antibodies are not made in most people because a substantial fraction of H structures are converted to A or B determinants.

IgM isoagglutinins efficiently trigger the complement cascade and they circulate in human plasma at titers sufficient to cause complement-dependent lysis of transfused erythrocytes that display the corresponding blood group antigens. Such rapid red cell lysis is associated with an immediate, or acute, transfusion reaction, which can lead to hypotension, shock, acute renal failure, and death from circulatory collapse. This problem is avoided by ensuring that the ABO type of transfused red cells is compatible with the recipient’s ABO type. Thus, an A recipient may receive red cells from another A person or from an O person, but not from a person of type B or AB. Blood banks perform typing and cross-matching assays. First, units of red cell products typed for the A and B antigens are chosen to match the patient’s ABO type. To ensure that these are truly “compatible,” the patient’s serum is cross-matched by mixing with a small aliquot of each prospective red cell unit. Red cells of compatible units do not agglutinate (form a clump), whereas incompatibility is indicated by agglutinated red cells formed by antibodies in the patient’s serum. Blood typing is used to ensure compatibility not only for red blood cell transfusions, but also for transfusion with plasma. Similar ABO compatibility concerns are important in heart, kidney, liver, and bone marrow transplantation procedures. The “type and cross” procedures have virtually eliminated ABO blood group transfusion reactions in the developed world. In the rare instances where a transfusion reaction occurs, the cause is usually human error.

Cross-matching procedures helped to identify a rare ABO blood group phenotype termed the Bombay phenotype, so named because the first identified individual lived in that city. Affected persons have red cells deficient in H, A, and B antigens, whereas their sera contain IgM antibodies that react with red cells from virtually all donors, including O red cells (H antigen-positive, A and B antigen-negative). Bombay people have inactive H and Se genes and are therefore incapable of synthesizing A, B, or H determinants in any tissue because of the absence of both α1–2FucT enzymes. They exhibit robust titers of anti-H, anti-A, and anti-B IgM antibodies and are therefore incompatible with red cells of all donors except those of the same Bombay (H-deficient) blood type. A related phenotype, termed para-Bombay, occurs in people with an inactive H gene but at least one functional Se allele (secretor-positive).

The fact that Bombay individuals appear healthy implies that developmental or physiological functions for the A, B, and H antigens, if they ever existed, are no longer relevant. It has been proposed that polymorphisms at the ABO locus may provide a selective advantage for protection from certain infectious agents, and recent genetic evidence tends to support this contention. Such selective pressures might therefore have maintained population polymorphisms at the ABO locus. In fact, a variety of associations have been made between the ABO blood group phenotype and relative risk for a spectrum of diseases.

One possible association concerns the well-known correlation between ABO status and plasma levels of von Willebrand factor (VWF), which is a glycoprotein involved in hemostasis. In the mouse, the circulating half-life of VWF (and thus its level in plasma) varies according to the nature of the alleles at a locus that encodes a β1–4GalNAcT, which modifies VWF glycans. A low serum VWF level is associated with an allele that directs expression of the β1–4GalNAcT to vascular endothelial cells, a major site of VWF synthesis. Glycoforms of VWF that carry the β1–4GalNAc modification are rapidly removed from plasma by the asialoglycoprotein receptor in liver. In contrast, mice with normal VWF levels do not express the β1–4GalNAcT in vascular endothelium, so their VWF is cleared less rapidly from the circulation. In humans, a similar mechanism could also account for the generally lower VWF levels in people with the O blood group, since VWF carries A and B blood group determinants in humans, and these structures may be recognized with lower affinities by the asialoglycoprotein receptor.

A second possible association is the proposed role for gastrointestinal ABO status and Lewis blood group antigens in the pathogenesis of the ulcerogenic spirochete Helicobacter pylori. The A blood group is associated with a modest increase in the risk of stomach cancer, whereas blood group O is associated with a slight increase in the risk of developing peptic ulcers. Although both of these disorders are now clearly associated with infection by H. pylori, it remains to be determined if there is a mechanistic relationship between ABO status and the consequences of H. pylori infection. A third explanation is that enveloped viruses carry the ABH glycans of their hosts and are thus susceptible to lysis upon first encountering the body fluids of another individual with an ABO-incompatible type. Finally, differences in susceptibility to severe complications of malaria appear to be affected by ABO blood groups. Several or all of these mechanisms may explain why the ABH system has remained active for more than 50 million years of primate evolution.


The Lewis blood group antigens are a related set of glycans that carry α1–3/α1–4 fucose residues (Figure 13.10). The term Lewis derives from a family who suffered from a red blood cell incompatibility that helped lead to the discovery of this blood group. The Lewisa antigen (Lea) is synthesized by an α1–3/α1–4FucT encoded by the Lewis (Le) blood group locus (Figure 13.11). The Lewisb antigen (Leb) is synthesized by the concerted actions of the Lea α1–3/α1–4FucT and the α1–2FucT encoded by the Se gene (Figure 13.11). The nature of the alleles at the Le and Se loci determines the complement of Lewis glycans that are synthesized by an individual. Secretor-positive individuals convert type-1 units to type-1 H determinants that may be acted on by the Lewis α1–3/α1–4FucT to form the Leb determinant (Figure 13.11). Nonsecretors who do not synthesize type-1 H determinants in secretory epithelia express the Lea determinant through the action of the Lewis α1–3/α1–4FucT (Figure 13.11). Individuals with an inactive Le locus are termed Lewis-negative. Lewis-negative secretors express type-I H determinants that cannot be converted to Lea or Leb determinants (Figure 13.11). Lewis-negative nonsecretors express type-1 units that are devoid of fucose.

FIGURE 13.10. Type-1 and -2 Lewis determinants.

FIGURE 13.10

Type-1 and -2 Lewis determinants. Type-1 and -2 units differ in the linkage of the outermost galactose (β1–3 or β1–4, respectively) and in the linkage of fucose to the internal GlcNAc (α1–4 or α1–3, (more...)

FIGURE 13.11. Lewis blood groups.

FIGURE 13.11

Lewis blood groups. The type-1 Lewis blood group determinants on glycoproteins and glycolipids (R) are characterized by the presence or absence of the Secretor locus α1–2FucT and the Lewis locus α1–3/α1–4FucT. (more...)

Expression of Lea and Leb glycans and the Lewis α1–3/α1–4FucT is largely restricted to the same epithelia that express the Se α1–2FucT. Thus, soluble forms of these antigens are released into secretions and body fluids. Lea and Leb antigens are also detectable on red cells. However, the precursors of red cells do not synthesize these determinants. Instead, Lewis antigens are acquired by the red cell membrane through passive adsorption of Lewis-positive glycolipids that circulate in plasma as lipoprotein complexes and aqueous dispersions. Antibodies against the Lea antigens have been implicated in occasional transfusion reactions.

Other members of the Lewis blood group family include the Lewisx (Lex) and Lewisy (Ley) determinants and forms of the Lea and Lex determinants that are sialylated and/or sulfated (Figure 13.10). These structures are formed through the actions of one or more α1–3FucTs that are distinct from the Lewis α1–3/α1–4FucT. Some members of the Lewis blood group antigen family have important functions in selectin-dependent leukocyte and tumor cell adhesion processes. Most strongly implicated are the sialylated and/or sulfated determinants represented by sialyl Lex and its sulfated variants (Figure 13.10), which function as selectin ligands on glycoproteins and glycolipids of leukocytes and probably also on tumor cells (Chapters 31 and 44).

The Lewis blood group antigens have also been proposed to function in the pathogenesis of H. pylori, the causative agent in chronic active gastritis and which is associated with hypertrophic gastropathy, duodenal ulcer, gastric adenocarcinoma, and gastrointestinal lymphoma (see Chapter 34). This organism adheres to the gastric mucosa that express Lewis blood group antigens. H. pylori itself also expresses Lewis antigens in a case of molecular mimicry. Antibodies generated against the bacterial Lewis antigens have been found in humans and may function to enhance colonization by H. pylori. However, the physiological relationship between H. pylori pathobiology and Lewis or ABO blood group status remains to be established.


The P blood group antigens are expressed on glycolipids of the red cell membrane and in tissues, including the urothelium (Figure 13.12). Little is known about the genes or glycosyltransferases that direct the expression of P blood group antigens. The synthesis of P antigens involves two different pathways, each of which begins with lactosylceramide as a common precursor (Figure 13.12). In one pathway, P antigen biosynthesis is initiated by an α1–4GalT (Pk transferase) that synthesizes the Pk antigen. The Pk antigen may be modified by a β1–3GalNAcT (P transferase) to form the P antigen. In the second pathway, P1-antigen biosynthesis involves three sequential glycosylation reactions, again starting with lactosylceramide. The first two reactions lead to paragloboside synthesis. Paragloboside is a substrate of the P1 transferase, which forms the P1 antigen. Expression of these antigens is polymorphic in humans. The most common P blood group is P1. These individuals express the enzymes of both pathways and their red cells express both P and P1 antigens. P1 individuals also express small amounts of Pk because the P transferase does not completely convert all Pk into P determinants. The other common blood group phenotype is P2 in individuals with an inactive P1 transferase. Red cells from P2 individuals express normal levels of P and Pk antigens but are deficient in P1 determinants. Antibodies directed against P blood group antigens have been implicated in transfusion reactions in individuals with the P phenotype, who often carry antibodies against P, P1, and Pk determinants. Complement-fixing, cold-reactive anti-P antibodies known as “Donath–Landsteiner” antibodies have been implicated in intravascular hemolysis observed in a syndrome called paroxysmal cold hemoglobinuria (see Chapter 43).

FIGURE 13.12. Biosynthesis of antigens of the P blood group system: Pk, P, and P1.

FIGURE 13.12

Biosynthesis of antigens of the P blood group system: Pk, P, and P1. Symbol Key: Image symbol_key_small.jpg

Physiological functions for the P blood group antigens are not known. However, a role in the pathogenesis of urinary tract infections is suggested because various uropathogenic strains of Escherichia coli express adhesins that bind to the Galα1–4Gal moiety of the Pk and P1 antigens (see Chapter 34). The P1 determinant is expressed on the urothelium and may facilitate infection by mediating attachment of bacteria to the lining of the urinary tract. In fact, P1 individuals have a higher relative risk for urinary tract infections and pyelonephritis. In addition, the adhesion of a pyelonephritic strain of E. coli to renal tissue is mediated by a bacterial adhesin specific for the Galα1–4Gal structure, and deficiency of the adhesin severely attenuates the pyelonephritic activity of the organism.

The P blood group antigens may also have a role as receptors for the human parvovirus B19. This virus causes erythema infectiosum and leads to congenital anemia and hydrops fetalis following infection in utero. It is also associated with transient aplastic crisis in patients with hemolytic anemia and with cases of pure red cell aplasia and chronic anemia in immunocompromised individuals. Parvovirus B19 replication is restricted to erythroid progenitor cells. An adhesive interaction between the virion and P antigen–active glyco-lipids is involved in viral infection of erythroid progenitors. Individuals with the P blood group phenotype are apparently resistant to parvovirus B19 infection.


The Galα1–3Gal epitope is synthesized on type-2 units on glycolipids and glycoproteins by a specific α1–3GalT (Figure 13.13). This epitope and the α1–3GalT are expressed by New World primates and many nonprimate mammals, but they are absent from the cells and tissues of Old World primates including Homo sapiens. The molecular basis for the clade-specific absence of this enzyme is inactivation of the gene encoding the α1–3GalT in primate taxa. A physiological function for the Galα1–3Gal epitope has not been identified.

FIGURE 13.13. Structure and synthesis of the Galα1–3Gal antigen.

FIGURE 13.13

Structure and synthesis of the Galα1–3Gal antigen. The α1–3GalT uses unsubstituted type-2 units on glycoproteins or glycolipids (R) to form the Galα1–3Gal terminal epitope. Symbol Key: Image symbol_key_small.jpg

Species that do not express the Galα1–3Gal epitope, including humans, carry naturally occurring anti-Galα1–3Gal antibodies in their serum, likely because of immunization through exposure to microbial antigens similar or identical to the Galα1–3Gal epitope. Anti-Galα1–3Gal antibodies present a major barrier to the use of porcine and other non-primate organs for xenotransplantation in humans, because they bind to Galα1–3Gal epitopes on the vascular endothelium of the xenotransplants and cause hyperacute graft rejection through complement-mediated endothelial cell cytotoxicity. Efforts are in progress to overcome this barrier by using animal organ donors that have been genetically modified (see Chapter 38). Approaches include transgenic expression of enzymes such as the H α1–2FucT that diminishes Galα1–3Gal expression by diverting type-2 units toward H antigen synthesis. The creation of animal organ donors with an inactive α1–3GalT locus represents a definitive solution to the problem. Unfortunately, however, pig tissues lacking Galα1–3Gal elicit a graft rejection reaction to other pig antigens. Anti-Galα1–3Gal antibodies have also been shown to significantly diminish the infective efficiency of recombinant retroviruses because packaging cell lines used to propagate these viruses express the Galα1–3Gal epitope. As with the ABO antigens, this may reflect a natural mechanism that acts to restrict the interspecies spread of retroviral genomes. The problem has been solved through the generation of packaging cell lines that are deficient in α1–3GalT. Recombinant glycoproteins for therapeutic use in humans must also be prepared in cells that do not express α1–3GalT.


The Forssman antigen (also known as globopentosylceramide) is a glycolipid that contains terminal N-acetylgalactosamine (GalNAc) in α1–3 linkage to the terminal GlcNAc of globoside (Figure 13.14). The Forssman antigen is expressed during embryonic and adult stages in many mammals. Humans carry anti-Forssman antibodies in their serum, suggesting that we do not synthesize the Forssman antigen. Although such antibodies are not consistently present, they may contribute to the pathogenesis of Guillain–Barré syndrome by binding to glycolipid components of peripheral nerve myelin. Similarly, there is evidence that small amounts of Forssman antigen may be found on human gastrointestinal epithelium, in various human cultured cells, and in pulmonary and gastointestinal tract carcinomas. These conflicting observations may reflect different specificities of the anti-Forssman monoclonal antibodies and differences in epitope reactivities with respect to detection methods. The function of the Forssman antigen is not known. However, anti-Forssman antibodies can disrupt tight junction formation, apical-basal polarization, and cell adhesion, suggesting that this molecule may participate in cell–cell adhesion and communication processes.

FIGURE 13.14. Forssman antigen biosynthesis.

FIGURE 13.14

Forssman antigen biosynthesis. The glycolipid globoside serves as the substrate for the Forssman α1–3GalNAcT that forms globopentosylceramide, also termed the Forssman glycolipid. Symbol Key: Image symbol_key_small.jpg


Glycans with sulfated terminal β-linked GalNAc are found on the pituitary glycoprotein hormones lutropin (LH) and thyrotropin (TSH) but not on follicle-stimulating hormone (FSH), although it is made in the same cells. These heterodimeric glycoproteins contain a common α subunit and a unique β subunit. Each subunit carries biantennary N-glycans. The biantennary N-glycans on TSH and LH contain an unusual 4-O-sulfated GalNAc attached to one or both GlcNAc residues (Figure 13.15). This contrasts with the N-glycans on FSH and on most N-glycans, in which GlcNAc residues are substituted with β1–4 galactose, which is often extended by α2–3 or α2–6 sialic acid (see Chapter 8). A free α subunit common to LH, TSH, and FSH is present in pituitary cells and it also carries this determinant, as do other glycoproteins synthesized by the pituitary and elsewhere (e.g., on the O-glycans of proopiomelanocortin). Synthesis of the sulfated GalNAc determinant is controlled by a β1–4GalNAcT that acts on GlcNAc residues of biantennary N-glycans (Figure 13.15). The terminal β1–4GalNAc is then sulfated by a sulfotransferase also expressed in pituitary cells. In some tissues, including the pituitary, the β1–4GalNAc is subsequently modified by an α2–6 sialic acid. GlcNAc residues extended with a β1–4GalNAc may also be modified by an α1–3 fucose residue. Both β1–4GalNAcT and β1–4GalT enzymes are expressed in pituitary cells, and the N-glycans on LH and TSH carry the unusual β1–4GalNAc. However, N-glycans on the related glycoprotein FSH carry the more common β1–4Gal residue. This protein-specific glycosylation is a consequence of interactions between the β1–4GalNAcT and a specific peptide motif present on the combined αβ subunits of LH and TSH. This interaction causes an increase in the catalytic efficiency of the β1–4GalNAcT that modifies biantennary N-glycans on LH and TSH at the expense of the competing β1–4GalT, which does not interact with the peptide motif. In contrast, the peptide motif recognized by the β1–4GalNAcT is not present in the β subunit of FSH, and the recognition motif on the α subunit of FSH is not accessible to the enzyme. Consequently, the biantennary N-glycans on FSH are modified exclusively by the competing β1–4GalT.

FIGURE 13.15. Structure and synthesis of N-glycans bearing terminal N-acetylgalactosamine (GalNAc), including those with sulfated-GalNAc found on the pituitary hormones lutropin (LH) and thyrotropin (TSH).

FIGURE 13.15

Structure and synthesis of N-glycans bearing terminal N-acetylgalactosamine (GalNAc), including those with sulfated-GalNAc found on the pituitary hormones lutropin (LH) and thyrotropin (TSH). Symbol Key: Image symbol_key_small.jpg

These differential glycosylation events have profound consequences for the ovulatory cycle in vertebrates. Circulating LH levels rise and fall in a highly pulsatile manner. This assures maximal stimulation of the ovarian LH receptor at the preovulatory surge, since sustained high LH levels would lead to LH receptor desensitization. The rise and fall in LH levels is due in part to pulsatile release of the hormone by the pituitary. However, it is also the consequence of rapid clearance of LH from the circulation mediated by a receptor that binds the terminal sulfated-GalNAcβ1–4GlcNAc determinant. This receptor is expressed in the liver by hepatic endothelial cells and by Kupffer cells. LH binding is followed by internalization and lysosomal degradation. The liver receptor is identical in protein sequence to that of the macrophage mannose receptor. However, in liver, the receptor recognizes sulfated-GalNAc via an R-type lectin domain, whereas in macrophages, the receptor recognizes mannose via an L-type lectin domain (see Chapter 28). An additional molecular basis for this difference is that in liver, the receptor is a dimer, whereas in macrophages, the receptor is a monomer. The receptor is now called the Man/GalNAc-SO4 receptor.

Similar GalNAcβ1–4GlcNAc (LacdiNAc) termini occur on N-glycans from other vertebrate sources, such as bovine milk, rat prolactin, and kidney epithelial cells, as well as in invertebrates such as snails. These residues generally do not become sulfated as in pituitary hormones, but they are frequently α2–6-sialylated in vertebrates. It is not yet clear how the GalNAcTs responsible for producing LacdiNAc are related to the enzyme that acts specifically on pituitary hormones.


The addition of GalNAc to glycans terminating with α2–3 sialic acid may occur on glycoproteins and glycolipids (Figure 13.16). On glycoproteins, this structure forms the human Sda blood group. In mouse, it was first described on mouse cytotoxic T lymphocytes (CTLs) and was termed the CT antigen. On glycolipids, this terminus forms the ganglioside GM2. The human Sda antigen was first identified in N-glycans of Tamm–Horsfall glycoprotein from human urine. The Sda determinant is formed by the addition of β1–4GalNAc to the Gal of α2–3-sialylated type-2 units (Figure 13.16). A β1–4GalNAcT capable of catalyzing this reaction exists in human kidney and urine, intestine, colon, and blood plasma. Both human and mouse β1–4GalNAcTs transfer GalNAc to N- and O-glycans present on glycoproteins but not to the glycolipid GM3 (Siaα2–3Galβ1–4Glc-Cer), even though both can efficiently use 3′-sialyllactose (Siaα2–3Galβ1–4Glc) as a substrate in vitro. In the mouse, the Sda antigen(s) is recognized by IgM monoclonal antibodies termed CT1 and CT2, which were isolated for their ability to block lysis of cellular targets by a murine CTL clone. The function of the Sda determinant in humans or mice is unknown. Rare humans lack the determinant and form naturally occurring antibodies against it, but they exhibit no apparent pathophysiology. As noted earlier, rare strains of mice with a dominantly inherited form of von Willebrand’s disease express the β1–4GalNAcT aberrantly in vascular endothelium. Since the blood-clotting protein VWF is also expressed by endothelial cells, the presence of the β1–4GalNAcT in this unusual location generates VWF carrying the Sda determinant. This VWF glycoform is rapidly cleared from the circulation, accounting for the low levels of VWF circulating in such strains of mice. Rapid clearance is mediated by the asialoglycoprotein receptor in liver, which exhibits affinity for terminal GalNAc.

FIGURE 13.16. Synthesis of the human Sda or mouse CT antigen and the glycolipid GM2.

FIGURE 13.16

Synthesis of the human Sda or mouse CT antigen and the glycolipid GM2. Symbol Key: Image symbol_key_small.jpg

The glycolipid form of the Sda determinant, termed GM2, is widely expressed in the central and peripheral nervous systems and in the adrenal gland, suggesting potential biological roles in these organs. In fact, mice homozygous for a null mutation at the GM2 synthase locus exhibit modest conduction defects in the peripheral nervous system and male sterility (see Chapter 38).


Sialic acid in α2–3 linkage is found on N-glycans, O-glycans, and glycolipids. A family of at least six different α2–3 sialyltransferases (ST3Gal-I to ST3Gal-VI) are responsible for its synthesis (Figure 13.17). ST3Gal-III and ST3Gal-IV are expressed in most cells of adult mammals. In contrast, ST3Gal-I transcripts in humans and mice are abundant in the spleen, liver, bone marrow, thymus, and salivary glands and are less abundant in other tissues. ST3Gal-II is also more restricted in expression pattern with transcripts most abundant in the brain, where α2–3-sialylated glycolipids are abundant (Figure 13.17). ST3Gal-V is expressed in the brain, skeletal muscle, testes, and liver. In vertebrates, α2–3 sialic acid residues are found on terminal galactose (Figure 13.17). Glycans terminating with galactose are substrates for the transfer of α1–3 fucose (Figure 13.10 and Chapter 31), β1–4GalNAc in limited circumstances (Figure 13.15), α2–6 sialic acid (Figure 13.18), α2–8 sialic acid (Figure 13.19 and Figure 13.20), and SO4 (Figure 13.10). In contrast, the α 2–3-sialylated glycans presented in Figure 13.17 are generally not substrates for other enzymes, including α1–2FucTs, α1–3GalT, GlcNAcTs, or GalNAcTs. The latter enzymes compete with terminal α2–3 sialyltransferases. Although most α2–3 sialic acid on glycoproteins is found on complex N-glycans (see Chapter 8) and O-GalNAc glycans (see Chapter 9), it also occurs on O-fucose and O-mannose glycans that are found on a limited subset of glycoproteins (see Chapter 12). As discussed in Chapter 31, the ligands recognized by the selectin family of leukocyte adhesion molecules include α2–3-sialylated glycans, modified additionally by α1–3 fucose or by α1–3 fucose and sulfate. Glycans bearing α2–3 sialic acid may contribute to the circulating half-life of plasma glycoproteins by virtue of “masking” terminal galactose residues that would contribute to the removal of glycoproteins from serum by the asialoglycoprotein receptor (see Chapter 31). However, mice deficient in this receptor do not have increased plasma levels of desialylated glycoproteins or lipoproteins in their circulation. In fact, recent studies demonstrate that the asialoglycoprotein receptor can also recognize glycans terminating in α2–6 sialic acid attached to GalNAcβ1–4 or Galβ1–4 (Figure 13.15)

FIGURE 13.17. Synthesis of glycoproteins and glycolipids bearing terminal α2–3 sialic acid transferred by the ST3Gal family of sialyltransferases.

FIGURE 13.17

Synthesis of glycoproteins and glycolipids bearing terminal α2–3 sialic acid transferred by the ST3Gal family of sialyltransferases. Enzymes in parentheses contribute at relatively low levels in vitro to the reactions indicated. Symbol (more...)

FIGURE 13.18. Synthesis of α2–6 sialic acid on O-glycans and glycolipids (see Chapters 9 and 10) by the ST6GalNAc family of sialyltransferases.

FIGURE 13.18

Synthesis of α2–6 sialic acid on O-glycans and glycolipids (see Chapters 9 and 10) by the ST6GalNAc family of sialyltransferases. Enzymes in parentheses contribute at relatively low levels in vitro to the reactions indicated. Symbol Key: (more...)

FIGURE 13.19. Structure and synthesis of polysialic acid on N-glycans.

FIGURE 13.19

Structure and synthesis of polysialic acid on N-glycans. Symbol Key: Image symbol_key_small.jpg

FIGURE 13.20. Structure and synthesis of polysialic acid on glycolipids.

FIGURE 13.20

Structure and synthesis of polysialic acid on glycolipids. Enzymes in parentheses contribute at low levels in vitro to the reactions indicated. Symbol Key: Image symbol_key_small.jpg

In other studies, the role of ST3Gal-I in the production of the Siaα2–3Galβ1–3GalNAcα-Ser/Thr glycan is important for the viability of peripheral CD8+ T cells. Mice lacking ST3Gal-I exhibit decreased cytotoxic T cell responses with an increase in the apoptotic death of naïve CD8+ T cells. The timing, specificity, and location of this enhanced apoptosis indicate an endogenous lectin, yet to be recognized, that binds to Galβ1–3GalNAcα-Ser/Thr glycans (see Chapter 33).

Sialic acid in α2–3 linkage is recognized by the hemagglutinin in the envelope of influenza viruses from birds and pigs. Binding to sialic acid and subsequent release by neuraminidase are important for infection by influenza virus. Human influenza viruses bind more commonly to sialic acid in α2–6 linkage. Mutations in the hemagglutinin genes of influenza viruses from birds may lead to a human influenza pandemic partly because of alterations in the ability to infect human cells through hemagglutinin recognition of α2–6-linked sialic acid (see Chapter 39).

Structures bearing α2–3 sialic acid have also been implicated in bacterial pathogenesis. For example, glycans terminating in α2–3 sialic acid support the adhesion of H. pylori, which causes gastritis, gastric ulcers, and lymphoma of the gastrointestinal tract mucosa. The ganglioside GM1 (Galβ1–3GlcNAcβ1–4[Siaα2–3]Galβ1–4GlcβCer) is a receptor for cholera toxin produced by Vibrio cholerae and the heatlabile enterotoxin (LT-1) produced by enterotoxigenic E. coli (see Chapter 39). Glycan-based inhibitors are currently under evaluation in humans for their ability to diminish the symptoms and progression of cholera.


Sialic acid in α2–6 linkage is found on N-glycans, O-glycans, and glycolipids. A family of at least six different α2–6 sialyltransferases (Figure 13.18) (including ST6Gal-I, ST6Gal-II, and ST6GalNAc-I to ST6GalNAc-IV) catalyzes its transfer. In vertebrates, α2–6 sialic acid is found on terminal galactose, on terminal or subterminal GlcNAc, or, in the case of reactions catalyzed by ST6GalNAc-III, on an internal GalNAc. α2–6 sialic acid appears less ubiquitously on glycans compared to α2–3 sialic acid. Glycans with terminal α2–6 sialic acid are generally not modified further, except possibly by some members of the poly α2–8 sialyltransferase family, as discussed below. The products of ST6Gal-I are typically found on N-glycans, although in vitro assays indicate that β1–4 galactose on O-GalNAc glycans can be modified by this enzyme. ST6Gal-I is expressed at a relatively high level in hepatocytes and lymphocytes and is responsible for α2–6 sialylation of serum glycoproteins and glycoproteins of the antigen receptor complex in lymphocytes. In contrast, the α2–6-sialylated glycans from ST6GalNAc-I and ST6GalNAc-II are restricted to O-glycans. ST6GalNAc-III is responsible for transferring α2–6 sialic acid to the GalNAcα-Ser/Thr core of O-glycans, whereas ST6GalNAc-IV appears to use glycolipids as preferred acceptors.

As noted above (and in Chapter 39), α2–6 sialic acid serves as a receptor for many strains of influenza virus infectious for humans, and glycoproteins bearing α2–6 sialic acid are recognized and cleared from the circulation by asialoglycoprotein receptor.

Mice lacking the enzyme ST6Gal-I exhibit an immunodeficiency characterized by a diminished antibody response to T-lymphocyte-dependent and -independent antigens, reduced B-lymphocyte proliferation in response to cross-linking of the B cell surface glycoproteins CD40 and surface IgM, reductions in the expression of B cell surface IgM and CD22, and an approximately 65% reduction in serum IgM levels. B lymphocyte antigen receptor–dependent signal transduction processes are also attenuated in these mice. These observations disclosed an important role for ST6Gal-I in the immune system and indicated defects in B cell receptor (BCR) signaling (see Chapter 32). The extracellular domain of CD22 on B lymphocytes specifically recognizes Siaα2–6Galβ1–4GlcNAc-. In the absence of α2–6 sialic acid, CD22 exhibits increased clustering with the BCR and thus BCR signaling is dysregulated. Interestingly, mice lacking both ST6Gal-I and CD22 are not immunodeficient, suggesting that α2–6 sialic acid is a functional ligand for CD22.


Glycans modified by α2–8 sialic acid are developmentally regulated in the vertebrate central nervous system. α2–8-Sialylated glycans may also be expressed in nonneuronal tissues during embryogenesis and in the adult vertebrate, and these glycans have been observed on transformed cells. At least five different α2–8 sialyltransferases direct the synthesis of α2–8-sialylated glycans (Figure 13.19 and Figure 13.20). Two of these enzymes, termed ST8Sia-II (also called STX) and ST8Sia-IV (also called PST-1 or PST) catalyze the synthesis of linear polymers of 200 or more α2–8 sialic acid residues to give polysialic acid (PSA) (Figure 13.19). N-glycans with terminal α2–3 sialic acid (and possibly α2–6 sialic acid) serve as the substrate for attachment of the initial α2–8 sialic acid. This “initiation” reaction is followed by an “elongation” reaction in which each α2–8 sialic acid serves as the attachment site for the next α2–8 sialic acid. Both activities are performed by the ST8Sia-II and ST8Sia-IV enzymes.

PSA has been most extensively studied as a component of the neural cell-adhesion molecule (NCAM), a member of the immunoglobulin superfamily (see Chapter 14). The α subunit of the voltage-gated sodium channel may also carry PSA. Both ST8Sia-II and ST8Sia-IV are autocatalytic and synthesize PSA on their own N-glycans, although polysialylation is not a prerequisite for their polysialyltransferase activity. Autocatalysis may yield membrane-associated, poly α2–8-sialylated forms of the enzymes, which may be why some cultured cell lines that do not express NCAM or the sodium channel express surface PSA when transfected with ST8Sia-II or with ST8Sia-IV.

Regulated expression of PSA during development is directed in part by tissue-specific regulation of the expression of the ST8Sia-II and ST8Sia-IV genes (see Chapter 38). ST8Sia-II is prominently expressed in fetal brain, but it declines markedly during the post-natal period. In contrast, ST8Sia-IV continues to be expressed postnatally in the brain and in nonneuronal tissues such as the heart, lung, and spleen. Posttranscriptional mechanisms may also regulate PSA expression. PSA negatively modulates the homotypic adhesive properties of NCAM on opposing cells. The embryonic form of NCAM is extensively modified by PSA and is less able to participate in homotypic adhesive interactions than is the adult form of NCAM, which is not highly modified by PSA. PSA can also diminish interactions promoted by other adhesion molecules, including L1-dependent attachment to laminin or collagen. Since PSA is highly negatively charged, highly hydrated, and contributes up to one third of the molecular mass of NCAM, it is believed that PSA negatively modulates cell-adhesion processes by physical interference with apposition of the plasma membranes of adjacent cells. Mice lacking ST8Sia-IV exhibit reduced PSA in certain brain regions and have altered neuronal responses in the hippocampal CA1 region. Mice lacking ST8Sia-II have a distinct neuronal phenotype because of misguided migration of a subset of hippocampal neurons and ectopic synapses. When both ST8Sia-II and ST8Sia-IV are inactivated, mice lack PSA in the brain, have severe neuronal and other problems, and die precociously. However, this phenotype is reversed by also removing NCAM in a triple-gene-knockout strain. This shows that the loss of PSA from NCAM is the basis of the neuronal defects in the ST8Sia-II and ST8Sia-IV mutant mice and that the presence of NCAM lacking PSA causes the severe defects in double-knockout mice (see Chapter 38).

Certain glycolipids also carry α2–8 sialic acid linkages, which are constructed by three α2–8 sialyltransferases termed ST8Sia-I (also known as GD3 synthase), ST8Sia-III, and ST8Sia-V (Figure 13.20). They generate single or oligomeric α2–8 sialic acid but not the polymeric PSA synthesized by ST8Sia-II or ST8Sia-IV. The three enzymes are generally thought to act primarily on glycolipid substrates, but in vitro studies suggest that ST8Sia-III can also utilize N-glycans. The nature of products formed by these enzymes in vivo is not clear, but their in vitro substrate specificities are presented in Figure 13.20. These α2–8 sialyltransferases are expressed in the brain, where each exhibits a distinct developmentally regulated expression pattern. ST8Sia-I is also expressed in kidney and thymus. In vitro experiments imply that certain α2–8-sialylated glycolipids may participate in signal transduction processes in neuronal cell types. Inactivation of the ST8Sia-I gene in the mouse causes alterations in sensory neuron responses to pain.


In principle, any free hydroxyl group on a monosaccharide has the potential to be modified by sulfation. However, in vertebrates, glycan sulfation is generally restricted to galactose, GlcNAc, glucuronic acid, and GalNAc at internal or terminal positions. The internally sulfated glycans represented by heparin, heparan sulfate, and chondroitin sulfate proteoglycans are discussed in Chapter 16. This chapter discusses the sulfated glycans that contribute to L-selectin ligands, the HNK-1 epitope, keratan sulfate, and some pituitary glycoprotein hormones as mentioned above.

Sulfated glycans on the specialized endothelium that lines the postcapillary venules in peripheral lymph nodes and other secondary lymphoid organs mediate lymphocyte homing to peripheral lymph nodes. L-selectin on lymphocytes binds to the high endothelial venules (HEV) through recognition of L-selectin ligands present on O-GalNAc glycans of HEV glycoproteins. Sulfated forms of α2–3-sialylated, α1–3-fucosylated glycans represented by the sialyl Lewisx determinant (Figure 13.10) provide an essential contribution to L-selectin recognition of these glycoproteins. Sulfation occurs at C-6 of the galactose and the GlcNAc, both of which contribute to L-selectin ligand activity. Mice lacking these two sulfotransferases exhibit almost no homing of lymphocytes to HEV. The biosynthesis of sulfated L-selectin ligands and the enzymes that participate in this process are discussed in Chapter 31.

The HNK-1 antigen is a terminally sulfated glycan that was first described on human natural killer cells, and it is also called CD57. The HNK-1 epitope is expressed by a variety of cell types in the vertebrate nervous system, where its cell type–specific expression patterns change during neural development (see Chapter 38). The HNK-1 determinant is composed of a 3-O-sulfated glucuronic acid (GlcA) attached in α1–3 linkage to a terminal galactose (Figure 13.21). This structure is synthesized by specific glucuronosyltransferases that act on terminal (poly)-N-acetyllactosamine units of N-glycans. Glucuronylation is followed by 3-O-sulfation of the GlcA by one or more specific sulfotransferases. The HNK-1 epitope has also been described on O-glycans of glycoproteins, on proteoglycans, and on glycolipids. Two different glucuronic acid transferases participate in HNK-1 GlcA addition: GlcAT-P and GlcAT-S. They have very different activities for glycoprotein or glycolipid substrates in vitro and thus may generate functionally different HNK-1 epitopes in vivo. The HNK-1 epitope is present on a variety of neuronal cell glycoproteins, including NCAM, contactin, myelin-associated glycoprotein, telencephalin, L1, and P0 (the major glycoprotein of peripheral nerve myelin). There is evidence that HNK-1 can function as a ligand for laminin, L-selectin, P-selectin, and a cerebellar adhesion protein termed amphoterin. HNK-1 has also been shown to mediate homotypic adhesive interactions involving P0. HNK-1-dependent adhesive interactions have been implicated in cell migration processes involving cell–cell and cell–matrix interactions and are proposed to participate in reinnervation of muscles by motor neurons.

FIGURE 13.21. Synthesis and structure of the HNK-1 epitope.

FIGURE 13.21

Synthesis and structure of the HNK-1 epitope. (GlucuronosylT) Glucuronosyltransferase; (SulfoT) sulfotransferase. Symbol Key: Image symbol_key_small.jpg

Keratan sulfate represents a prominent posttranslational modification of several extra-cellular matrix proteins (see Chapter 16). Keratan sulfate is a poly-N-acetyllactosamine chain attached to proteins via a typical N-glycan (KS type I) or a typical O-glycan (KS type II). It is believed that keratan sulfate is synthesized in a stepwise manner during poly-N-acetyllactosamine synthesis, in coordination with a pair of 6-O-sulfotransferase activities (Figure 13.22). A keratan-specific N-acetylglucosamine 6-O-sulfotransferase activity (CHST6) transfers sulfate to terminal GlcNAc residues and will not effectively modify internal GlcNAc nor GalNAc in chondroitin sulfate. Mutations in this gene have been associated with macular corneal dystrophy in humans. A second, distinct 6-O-sulfotransferase (CHST5) also catalyzes 6-O-sulfation of GlcNAc in keratan sulfate.

FIGURE 13.22. Synthesis of keratan sulfate.

FIGURE 13.22

Synthesis of keratan sulfate. Different 6-O-sulfotransferases (SulfoT) transfer sulfate to the C-6 of galactose and GlcNAc in poly-N-acetyllactosamine chains. Keratan sulfate is attached to proteins (R) via an N-glycan (KS type I) or an O-glycan (KS type (more...)

The galactose 6-O-sulfotransferase CHST3 efficiently sulfates galactose within keratan sulfate poly-N-acetyllactosamine units, but it is apparently more efficient when the galactose is adjacent to a 6-O-sulfated GlcNAc. These observations imply that sulfation of GlcNAc residues can only occur when terminal GlcNAc exists during synthesis and elongation of poly-N-acetyllactosamine, whereas sulfation of galactose may occur at any time during poly-N-acetyllactosamine synthesis.

Corneal keratan sulfate (KS I) apparently maintains the proper spatial organization of the type I collagen fibrils in the cornea that promote transparency of this structure. Alterations in the degree of sulfation of keratan sulfate are associated with corneal opacity in a disease known as macular corneal dystrophy. Alterations in the amount of sulfation of brain keratan sulfate have been found in the brains of Alzheimer’s disease patients.


  1. Blumenfeld OO, Patnaik SK. Allelic genes of blood group antigens: A source of human mutations and cSNPs documented in the Blood Group Antigen Gene Mutation Database. Hum Mutat. 2004. pp. 8–16.http://www​.ncbi.nlm.nih​.gov/gv/mhc/xslcgi.cgi?cmd=bgmut/home [PubMed: 14695527]
  2. Yamamoto F. ABO blood group system—ABH oligosaccharide antigens, anti-A and anti-B, A and B glycosyltransferases, and ABO genes. Immunohematol. 2004;20:3–22. [PubMed: 15373665]
Copyright © 2009, The Consortium of Glycobiology Editors, La Jolla, California.
Bookshelf ID: NBK1892PMID: 20301230


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