Your privacy, your choice

We use essential cookies to make sure the site can function. We also use optional cookies for advertising, personalisation of content, usage analysis, and social media.

By accepting optional cookies, you consent to the processing of your personal data - including transfers to third parties. Some third parties are outside of the European Economic Area, with varying standards of data protection.

See our privacy policy for more information on the use of your personal data.

for further information and to change your choices.
Skip to main content
Download PDF

Human Gb3/CD77 synthase: a glycosyltransferase at the crossroads of immunohematology, toxicology, and cancer research

Cellular & Molecular Biology Letters volume 29, Article number: 137 (2024) Cite this article

Abstract

Human Gb3/CD77 synthase (α1,4-galactosyltransferase, P1/Pk synthase, UDP-galactose: β-d-galactosyl-β1-R 4-α-d-galactosyltransferase, EC 2.4.1.228) forms Galα1 → 4Gal structures on glycosphingolipids and glycoproteins. These glycans are recognized by bacterial adhesins and toxins. Globotriaosylceramide (Gb3), the major product of Gb3/CD77 synthase, is a glycosphingolipid located predominantly in plasma membrane lipid rafts, where it serves as a main receptor for Shiga toxins released by enterohemorrhagic Escherichia coli and Shigella dysenteriae of serotype 1. On the other hand, accumulation of glycans formed by Gb3/CD77 synthase contributes to the symptoms of Anderson–Fabry disease caused by α-galactosidase A deficiency. Moreover, variation in Gb3/CD77 synthase expression and activity underlies the P1PK histo-blood group system. Glycosphingolipids synthesized by the enzyme are overproduced in colorectal, gastric, pancreatic, and ovarian cancer, and elevated Gb3 biosynthesis is associated with cancer cell chemo- and radioresistance. Furthermore, Gb3/CD77 synthase acts as a key glycosyltransferase modulating ovarian cancer cell plasticity. Here, we describe the role of human Gb3/CD77 synthase and its products in the P1PK histo-blood group system, Anderson–Fabry disease, and bacterial infections. Additionally, we provide an overview of emerging evidence that Gb3/CD77 synthase and its glycosphingolipid products are involved in cancer metastasis and chemoresistance.

Graphical Abstract

Introduction

Human Gb3/CD77 synthase (α1,4-galactosyltransferase, UDP-galactose: β-d-galactosyl-β1-R 4-α-d-galactosyltransferase), known among immunohematologists as P1/Pk synthase, is encoded in humans by the A4GALT gene located on chromosome 22q13.2. It catalyzes the transfer of galactose residue from UDP-Gal to galactose-terminated glycosphingolipid (GSL) and glycoprotein acceptors. The many names of this enzyme reflect distinct fields in which its activity has been found relevant, such as glycobiology, immunohematology, toxicology, and oncology. Similarly, globotriaosylceramide, commonly referred to as Gb3, is also called CD77, the Pk antigen, or ceramide trihexoside (CTH). This may lead to misunderstandings and inaccessibility of reported data among researchers accustomed to the use of other terms.

In this review, we aim to summarize and present the most important data on the functions of Gb3/CD77 synthase and its products from different perspectives. On the one hand, glycans produced by Gb3/CD77 synthase determine phenotypes of the P1PK blood group system. On the other hand, we aim to emphasize that they are histo-blood group antigens present on many cell types other than erythrocytes, such as lymphocytes, renal endo- and epithelium, epithelium of gastrointestinal tract, or neurons. Hence, the role of these glycans as receptors for toxins and lectins released by such deadly pathogens as Shiga toxin-producing Escherichia coli (STEC), Staphylococcus aureus, or Pseudomonas aeruginosa is of particular significance. Moreover, since the products of Gb3/CD77 synthase are overexpressed in human colorectal, gastric, pancreatic, and high-grade serous ovarian cancer, they have diagnostic and therapeutic potential.

Structure and localization of Gb3/CD77 synthase

Classified under the number EC 2.4.1.228, human Gb3/CD77 synthase belongs to the 32nd family of glycosyltransferases (GT32; Carbohydrate-Active enZYmes database, www.cazy.org). In silico predictions place the enzyme among type II transmembrane proteins belonging to a GT-A superfamily. Interestingly, the Gb3/CD77 synthase amino acid sequence shows the closest similarity to α1,4-N-acetylglucosaminyltransferase, encoded by A4GNT (Łukasz Sobala, personal communication).

The polypeptide chain of Gb3/CD77 synthase comprises 353 amino acids, of which residues 1–22 probably represent the cytoplasmic tail, 23–43 belong to the transmembrane domain, and 44–353 form the stem region and catalytic domain facing Golgi lumen (Fig. 1A). The catalytic domain contains a DxD motif (D192TD according to UniProt Q9NPC4) (Fig. 1B), which is indispensable for the enzymatic activity of Gb3/CD77 synthase, presumably through establishing interactions with divalent ions, such as Mn2+, at the catalytic center [1, 2]. The enzyme has two N-glycosylation sites, N121 and N203 (Fig. 1B), and both are occupied. However, while glycosylation of the N121 site seems dispensable, the N-glycan occupying N203 is crucial for Gb3/CD77 synthase localization and activity [3, 4].

Fig. 1
figure 1

Structure of Gb3/CD77 synthase. A The 3D structure model predicted by AlphaFold (https://alphafold.ebi.ac.uk/entry/Q9NPC4); amino acid residues 1–22 compose the cytoplasmic tail, 23–43 form the transmembrane domain, and 44–353 encompass the stem and catalytic domain facing Golgi lumen. B A schematic representation of the Gb3/CD77 synthase polypeptide chain with important peptide motifs highlighted

Full size image

Gb3/CD77 synthase resides in the trans-Golgi network (TGN) [3, 5, 6] where it localizes owing to the interactions with COPI vesicle-associated proteins, Golgi phosphoprotein 3 (GOLPH3) [7] and transmembrane 9 superfamily 2 protein (TM9SF2) [5, 8]. COPI-dependent retrograde transport of Golgi-resident enzymes is a crucial mechanism of the Golgi cisternal maturation model [9]. In particular, GOLPH3 and its paralog GOLPH3L control the recruitment of Golgi-resident enzymes in trans cisternae and loading into COPI-dependent retrograde vesicles [10]. Gb3/CD77 synthase is recognized by GOLPH3 through an N-terminal LxxR motif (L7LLR according to UniProt Q9NPC4), similarly to the lactosylceramide synthase (B3GNT5) and several other enzymes engaged in GSL biosynthesis [7]. Moreover, the activity of Gb3/CD77 synthase depends on interactions with lysosome-associated protein transmembrane 4α (LAPTM4A) [5, 8].

α1,4-Galactosyltransferase activity

Human Gb3/CD77 synthase catalyzes the transfer of galactose from UDP-Gal to galactose-terminated GSLs and N-glycan chains, creating Galα1 → 4Gal structures. The consensus human α1,4-galactosyltransferase can use several GSLs as acceptors, including galactosylceramide, lactosylceramide, and nLc4/paragloboside, giving rise to galabiosylceramide (Ga2, Gal2Cer), globotriaosylceramide (Gb3, CD77, the Pk antigen, CTH), and the P1 antigen (nLc5), respectively (Fig. 2) [11, 12]. Galabiosylceramide is the precursor of the gala-series, Gb3 initiates the globo-series, whereas P1 belongs to the neolacto-series of GSLs. Human Gb3/CD77 synthase was long believed to use only GSLs as acceptors. Recently, we challenged that paradigm and demonstrated that the enzyme attaches galactose residues also to N-glycans on glycoproteins [13].

Fig. 2
figure 2

A schematic representation of the biosynthesis of GSLs and N-glycans produced by Gb3/CD77 synthase. Galα1 → 4Gal structures are highlighted in green, and Galα1 → 4GalNAc in red. Color coding according to the Symbol Nomenclature for Glycans (SNFG) [120]

Full size image

Most probably, Gb3/CD77 synthase also produces P1 glycotopes (Galα1 → 4Galβ1 → 4GlcNAc) carried by long GSLs belonging to the neolacto-series: heptaosylceramide and branched decaosylceramide containing two P1 trisaccharide units. These newly identified GSL species have been recently isolated from human gastric adenocarcinoma [14].

Astoundingly, a single point mutation (c.631C > G; rs397514502) (Fig. 3) resulting in the substitution of glutamine with glutamic acid at the 211 position of the enzyme polypeptide chain renders Gb3/CD77 synthase able to use N-acetylgalactosamine-terminated GSLs as acceptor substrates and synthesize the NOR antigen [12, 15]. Gb3/CD77 synthase p.Q211E (also called the NOR synthase) galactosylates globotetraosylceramide (Gb4, globoside), producing NOR1 (terminated with one Galα1 → 4GalNAc moiety [16]), which is subsequently elongated by the Gb4 synthase (β1,3-N-acetylgalactosaminyltransferase, B3GALNT1), forming NORint (Fig. 2). This GSL is further modified by the NOR synthase, leading to the formation of NOR2, encompassing two Galα1 → 4GalNAc repeats [15]. Thus, Gb3/CD77 synthase p.Q211E exhibits broadened acceptor specificity and, alongside Galα1 → 4Gal structures (on GSLs and glycoproteins), also synthesizes the Galα1 → 4GalNAc moiety of the glycosphingolipid NOR antigen. Modulation of glycosyltransferase acceptor specificity by a single amino acid substitution is unprecedented, and human Gb3/CD77 synthase remains its sole example.

Fig. 3
figure 3

Schematic representation of the A4GALT gene. The rectangles depict the three exons. The circles indicate the positions of SNPs associated with P1/P2 polymorphism (rs5751348 in red, rs2143918 and rs8138197 in blue), and the yellow circle represents the rs397514502 SNP resulting in p.Q211E substitution

Full size image

The presence of NOR antigen causes the rare NOR polyagglutination syndrome [17]. The first reported case was a resident of Norton, Virginia, hence the designation “NOR” [18]. Red blood cells (RBCs) of individuals expressing NOR were agglutinated by most ABO-compatible human sera owing to the commonly occurring natural anti-NOR antibodies [18,19,20]. It is noteworthy that the terminal Galα1 → 4GalNAc structure of NOR is extremely unique. It was never detected before in mammals, and among vertebrates, it was reported solely in Rana ridibunda egg jelly coat mucin [21]. However, a similar structure (Galα1 → [4GalNAcα1 → 4Galα1]n → 4GalNAc) was identified in galactosaminogalactan secreted by Aspergillus fumigatus [22]. Most human sera react with this polysaccharide, presumably owing to the presence of anti-NOR antibodies elicited by A. fumigatus infections.

A4GALT gene regulation and the genetic background of the P1PK histo-blood group system

Gb3, under the name of “Pk antigen,” along with P1 and NOR, are the three antigens constituting the P1PK histo-blood group system (International Society of Blood Transfusion (ISBT) no. 003). The P1PK system (originally designated as “P”) was discovered by Landsteiner and Levine in 1927, and since then has undergone numerous modifications following advances in understanding of its biochemical and genetic basis (reviewed in Refs. [23, 24]). Most individuals display P1/P2 phenotypes: P1 when the P1 antigen along with Pk (Gb3) is expressed on erythrocytes, and P2 when only Pk (Gb3) is present (Table 1). P1/P2 distribution varies greatly between populations; e.g., the incidence of the P1 phenotype ranges from 20–30% among Asians to 75–80% in Europeans [25].

Table 1 The phenotypes of the P1PK histo-blood group system and A4GALT genotypes (ψψ does not encode any functional protein)
Full size table

The gene encoding Gb3/CD77 synthase (A4GALT) encompasses three exons, and the entire open reading frame (ORF) is contained within exon 3 (Fig. 3). Various missense, nonsense, and frameshift mutations in the A4GALT ORF underlie the rare p phenotype (incidence estimated at 5.8 per million), found in individuals lacking any P1PK antigens [26]. In turn, the P1NOR phenotype, which is characterized by the presence of Pk (Gb3), P1, and NOR, originates from a single point mutation c.631C > G in the A4GALT ORF (rs397514502) causing the p.Q211E substitution [15]. The P1NOR phenotype has been identified in members of only two families to date, one living in the USA and one in Poland [18, 19].

The genetic basis of the P1/P2 phenotypic polymorphism posed a long-standing conundrum because it does not arise from mutations in the A4GALT ORF [27]. Since individuals with the P1 phenotype showed increased A4GALT transcript levels in comparison with P2, and it was generally accepted that the P1/P2 polymorphism is associated with transcriptional regulation of the A4GALT expression [28, 29]. Throughout the years, several single-nucleotide polymorphisms (SNPs) upstream from the coding region were proposed to determine the P1/P2 status [28,29,30], of which rs5751348 proposed by Lai and coworkers [30] proved to be the most important [31, 32]. rs5751348 is located in the A4GALT intron 1 (3084 bp downstream from the transcription initiation nucleotide of exon 1, Fig. 3), near rs2143918 and rs8138197, the SNPs usually linked with rs5751348 and hence also well associated with the P1/P2 polymorphism. rs5751348G (A4GALT P1 allele) sits within the binding motifs for transcription factors early growth response protein 1 (EGR1) and Runt-related transcription factor 1 (RUNX1) [33, 34]. EGR1 binds specifically to the P1 allele, thus activating its transcription [33]. RUNX1 was shown to bind the P1 allele-specific probes, and its knockdown resulted in downregulation of A4GALT expression [34].

Some individuals with the P2 phenotype produce anti-P1 antibodies, but they rarely cause hemolytic transfusion reactions [35, 36]. However, individuals with the null p phenotype develop so-called anti-PP1Pk (formerly known as “anti-Tja”) antibodies, which recognize Pk (Gb3) and P1, as well as the P antigen belonging to the GLOB histo-blood group system (ISBT no. 028). The P antigen is globotetraosylceramide (Gb4), produced directly from Pk (Gb3) (Fig. 2) and hence also absent in p individuals. Anti-PP1Pk antibodies can induce severe hemolytic transfusion reactions, recurrent spontaneous abortions, and sporadically mild hemolytic disease of the fetus and newborn [37]. Interestingly, most human sera contain antibodies that can recognize a purified Pk antigen (Gb3), but do not bind Pk-positive cells, nor do they agglutinate Pk-positive erythrocytes owing to the crypticity of cell-bound Pk [38, 39].

Tissue distribution of Gb3/CD77 synthase and its products

Expression of Gb3/CD77 synthase and its products is not restricted to the erythroid cell lineage. The A4GALT transcript is ubiquitously expressed in all human tissues, at high levels in the heart, kidney, spleen, liver, testis, and placenta and at lower levels in the lung, stomach, brain, colon, and muscle [11, 27].

Galabiosylceramide was detected in the kidney, urine, and cerebrospinal fluid [40,41,42]. In normal tissues, P1 antigen expression appears to be limited to erythrocytes [43]. Similarly, the P1 glycotope-capped N-glycoproteins have been found so far only on erythrocytes of P1P1 individuals [44].

However, Gb3 is widely distributed in human tissues. Its presence was reported in lymphocytes, platelets, heart, kidney, lung, liver, smooth muscle, epithelium of gastrointestinal tract, neurons, and central nervous system endothelium [43, 45,46,47,48]. Gb3 was detected in a subset of B lymphocytes in germinal centers, where it was suggested to play a role in the negative selection of B cells, hence being termed CD77 [49,50,51,52]. Recently, it was shown that Gb3 expressed on germinal center B cells promotes the translocation of CD19 to the B cell receptor complex and stimulates downstream signaling pathways [53]. Moreover, Gb3 is a major neutral GSL of T lymphocytes and macrophages [54]. In the kidney, Gb3 is expressed on endothelial as well as on epithelial tubular cells [55], where it presumably contributes to the reabsorption of low-molecular-weight proteins and albumin [56]. Moreover, Gb3 is also present in human milk and presumably may act as a decoy receptor for Shiga toxins, protecting breastfed infants from developing hemorrhagic colitis and hemolytic uremic syndrome [57, 58]. It should be noted that, in methods based on immunodetection on the cell surface, Gb3 is poorly accessible for monoclonal antibodies without protease pretreatment of the cells and hence its expression may be underestimated.

Gb3/CD77 synthase products as culprits of Anderson–Fabry disease

Under homeostatic conditions, the products of Gb3/CD77 synthase are degraded in lysosomes by α-galactosidase A (EC 3.2.1.22), encoded by the GLA gene located on chromosome Xq22.1. Mutations in the GLA gene causing α-galactosidase A deficiency underlie the lysosomal storage disorder Anderson–Fabry disease (OMIM #301500). In this disease, α-galactose-terminated GSLs accumulate in tissues, plasma, and urine. Gb3 and its deacetylated derivative lysoGb3 are the main accumulating GSLs and serve as disease biomarkers [59]. LysoGb3 most probably emerges as a product of acid ceramidase (N-acylsphingosine deacylase, EC 3.5.1.23), encoded by the ASAH1 gene, which can use Gb3 as a substrate when it is available in excess [60]. The GSL deposits also contain galabiosylceramide, the P1 antigen, and the B histo-blood group antigen [61]. The deposits are found in many tissues, particularly in the heart, kidney, peripheral nerves, eye, brain, skin, gastrointestinal tract, and auditory system [62]. In the kidney, GSLs accumulate in podocytes, glomerular endothelial, mesangial, and interstitial and distal tubular cells [63]. Interestingly, in a symptomatic mouse model of Fabry disease (generated by GLA gene knockout and human A4GALT overexpression), the highest accumulation of Gb3 was found in the spleen and liver, which do not display any abnormalities in the mouse model or patients [64]. Anderson–Fabry disease includes neurological, cardiac, renal, ocular, gastrointestinal, dermatological, and auditory symptoms, of which cardiac and renal dysfunction are the leading causes of death [65]. Approved treatment options involve enzyme replacement (agalsidase alfa and beta) and chaperone therapy (migalastat) [66]. Other agents for enzyme replacement therapy and for substrate reduction approach inhibiting glucosylceramide synthase (ceramide glucosyltransferase, UGCG) are also extensively studied. In addition, gene and messenger RNA (mRNA)-based therapies that focus on GLA are considered as a viable option. However, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated suppression of A4GALT seems to emerge as a novel substrate deprivation strategy [67, 68], and perhaps in the future small-molecule inhibitors of Gb3/CD77 synthase could be developed.

Gb3/CD77 synthase products as receptors for bacterial toxins and adhesins

Galα1 → 4Gal structures produced by Gb3/CD77 synthase are recognized by bacterial adhesins and toxins (reviewed in Refs. [45, 69, 70]). Gb3 is a crucial receptor for Shiga toxins (also known as “verotoxins” or “Shiga-like toxins”), which are major virulence factors released by Shiga toxin-producing Escherichia coli (STEC) and Shigella dysenteriae of serotype 1 [71]. Shiga toxins are composed of the catalytically active A subunit and a pentamer of B subunits that recognize cellular receptors; toxins with such structure are collectively referred to as AB5 class [72]. STEC infections can cause hemorrhagic colitis, which can deteriorate into hemolytic uremic syndrome, a life-threatening complication characterized by thrombocytopenia, anemia, and acute kidney failure [73]. Recently, it was shown that glycoproteins carrying P1 glycotopes can also serve as functional receptors for type 1 Shiga toxins [13, 74]. Galabiosylceramide and the GSL P1 antigen, as well as globotetraosylceramide (Gb4), showed Shiga toxin binding in early reports [75, 76]. However, recent studies corroborated only type 1 Shiga toxin interaction with P1, and its role in toxin internalization still requires elucidation [77].

Staphylococcal enterotoxin B (SEB) from Staphylococcus aureus is a major toxin responsible for food poisoning and toxic shock syndrome. It recognizes terminal Galα1 → 4Gal structures, preferring galabiosylceramide [78]. PapG adhesins, which are expressed at the top of uropathogenic E. coli (UPEC) P-fimbriae, bind both terminal and internal Galα1 → 4Gal structures [79, 80]. Similarly, SadP adhesin from Streptococcus suis, a zoonotic pathogen, which causes meningitis and septicemia, recognizes terminal and internal Galα1 → 4Gal structures present on GSLs and glycoproteins (P1 glycotopes carried by pigeon egg white ovomucoid) [81].

LecA lectin (also known as “PA-I” or “PA-IL”) from Pseudomonas aeruginosa, an opportunistic pathogen causing nosocomial infections, binds α-d-galactosylated glycans, including Gb3 and the P1 antigen, with a preference for Gb3 [82, 83]. A LecA homolog, PIIA lectin from Photorhabdus luminescens, which lives symbiotically in insect-infecting Heterorhabditis nematodes, also recognizes α-d-galactosylated glycans [84].

The role of Gb3/CD77 synthase and its products in cancer progression

Gb3 overexpression was discovered in Burkitt’s lymphomas, and Gb3 was designated as Burkitt’s lymphoma antigen [85, 86]. Later, increased Gb3 level was reported in colorectal cancer [87,88,89]. In a study by Kovbasnjuk and coworkers [87], Gb3 expression was linked to a metastatic and invasive phenotype; however, subsequent reports did not corroborate the correlation between Gb3 level and metastasis. Moreover, strong Gb3 expression was found in gastric adenocarcinoma in most tissues of intestinal and diffuse type [90]. Gb3 was also reported as one of the major GSLs in scirrhous gastric carcinoma tissues [91]. Increased Gb3 levels were detected in pancreatic cancer [89, 92]. Breast tumors of distinct clinicopathological types and their lymph node metastases showed increased Gb3 expression, predominantly in epithelial cancer cells [93,94,95]. Furthermore, different histological subtypes of epithelial ovarian cancer display Gb3 overexpression [96, 97]. Taking advantage of both Gb3 characteristics, as a tumor-associated antigen and Shiga toxin receptor, several therapeutic strategies are now being developed involving targeting cancer cells with Shiga toxin subunit B conjugated to chemotherapeutic drugs [98].

The P1 antigen has been found in serous ovarian cancer and endometrioid peritoneal cancer tissues, as well as on ovarian cancer cells IGROV-1, where it correlated with migratory potential [99]. Moreover, patients with tubal, peritoneal, and ovarian cancer show a decreased level of plasma-derived naturally circulating anti-P1 IgM antibody, and this observation can be applied in biochemical diagnosis of gynecological tumors [99]. Recently, P1 antigen expression was detected in pancreatic ductal and gastric adenocarcinoma [14, 100]. In the latter study, in addition to the classic pentaosylceramide P1, two novel GSL structures terminating with the P1 glycotope were identified: heptaosylceramide and a branched decaosylceramide carrying two P1 trisaccharide units.

Gb3/CD77 synthase initiates the biosynthesis of globosides (GSLs of the globo-series) and plays a particularly important role in the epithelial-to-mesenchymal transition (EMT) of cancer cells. EMT is often accompanied and even induced by changes in the GSL composition of the plasma membrane [97, 101]. It was found that the transcript level of the A4GALT gene (encoding Gb3/CD77 synthase) correlates negatively with mesenchymal characteristics in ovarian cancer. Its CRISPR/Cas9-mediated deletion triggers EMT and chemoresistance of ovarian cancer cells [2]. On the contrary, A4GALT knockdown by small interfering RNA (siRNA) decreased the migratory potential of colon cancer epithelial cells [87]. In pancreatic ductal adenocarcinoma cells, Gb3 expression was observed in the mesenchymal-like PaTu-8988T but not in the epithelial-like PaTu-8988S cell line [102].

Enhanced biosynthesis of GSLs originating from glucosylceramide (GlcCer) was demonstrated in many chemoresistant cancer cells [103]. In particular, Gb3 overexpression is associated with chemoresistance of several types of human cancer cells, such as ovarian, breast, and colon cancer [104,105,106]. Interestingly, the increased Gb3 level may induce radio- and chemoresistance of cancer cells through interactions with Hsp70 and cSrc, respectively, in plasma membrane lipid rafts [104, 107]. Cancer cells often show Hsp70 overexpression and abnormal localization in the plasma membrane, which contributes to their radioresistance [108]. It was also shown that interaction with Gb3 enables cancer-specific membrane localization of Hsp70 in colon and pancreatic cancers, as well as Burkitt’s lymphoma cells [107, 109, 110]. On the other hand, association of Gb3 and cSrc kinase in lipid rafts underlies the overexpression of p53 R273H mutant and chemoresistance of colon cancer cells [106]. Furthermore, interactions of Gb3 with cSrc kinase most probably cause overexpression of MDR1 (P-glycoprotein), triggering multidrug resistance in ovarian cancer cells [104]. Gb3 was also suggested to activate the Akt and ERK1/2 signaling pathways leading to MDR1 overexpression and resistance of breast cancer cells [105].

Conclusions and future perspectives

Human Gb3/CD77 synthase continues to astound and shift paradigms. Once considered an unremarkable single-product enzyme, it has emerged as a multifunctional glycosyltransferase, capable of synthesizing several different structures on GSLs and glycoproteins. While recent studies have significantly expanded our understanding of its activity, other yet-undiscovered glycoconjugates modified by Gb3/CD77 synthase may exist. Preliminary results obtained by Bereznicka and coworkers suggest that a recombinant soluble catalytic domain of Gb3/CD77 synthase can attach galactose residues to Galβ1 → 3GalNAc-terminated O-glycans, forming Galα1 → 4Galβ1 → 3GalNAc structures (Bereznicka et al. unpublished data). However, production of these structures in vivo is a matter of controversy, since O-glycans are readily capped by sialyltransferases with terminal Neu5Ac residues, thus presumably preventing elongation by Gb3/CD77 synthase.

The question that then arises is: why does human Gb3/CD77 synthase show a preference for GSL over glycoprotein acceptors (we estimate it forms 95% of Galα1 → 4Gal structures on GSLs)? The ABO histo-blood group system transferases, which are similarly capable of using both GSLs and glycoproteins as acceptors, synthesize the A and B blood group antigens primarily on glycoproteins (80% of ABH determinants on RBCs) and to a lesser extent on GSLs (20%) [111]. However, a recombinant soluble catalytic domain of Gb3/CD77 synthase efficiently galactosylates purified N-glycans and N-glycoproteins [13]. In the Golgi apparatus, glycosyltransferases are known to form homo- and heterooligomers, which significantly impacts their activity. Such multienzyme complexes are thought to act as assembly lines whereby the products of one reaction move along to become acceptors of the next reaction in the biosynthetic pathway [112, 113]. We presume that the low level of P1 glycotope biosynthesis on cellular glycoproteins may originate from preferential contribution of Gb3/CD77 synthase to multienzyme complexes responsible for GSL production in the Golgi. Recently, Mikolajczyk and coworkers provided evidence for the formation of Gb3/CD77 synthase homodimers as well as heterodimers with β1,4-galactosyltransferase 1 (B4GALT1) and β1,4-galactosyltransferase 5 (B4GALT5) [114]. Interestingly, Gb3/CD77 synthase showed preference for interactions with β1,4-galactosyltransferase 5, which is a GSL-specific enzyme, in contrast to glycoprotein-specific β1,4-galactosyltransferase 1.

The acceptor promiscuity of human Gb3/CD77 synthase, making it capable of synthesizing both Galα1 → 4Gal and Galα1 → 4GalNAc terminal structures, is a unique property among glycosyltransferases. As the crystal structure of Gb3/CD77 synthase remains to be solved, we can only speculate how p.Q211E substitution enables the enzyme to produce the Galα1 → 4GalNAc moiety of the NOR antigen. We suppose that this modification deepens the acceptor binding site in the enzyme catalytic center, enabling GalNAc-terminated GSL acceptors to fit in. Preliminary analysis of a Gb3/CD77 synthase model predicted by AlphaFold supports this hypothesis (Sobala et al., unpublished data).

The abundance of Gb3/CD77 synthase products in human tissues underscores their importance in bacterial infections, of which STEC seem to pose the greatest threat. The estimated number of acute illnesses worldwide caused by STEC infections is 2.8 million annually, and the disease is particularly dangerous for children and elderly persons [115]. Furthermore, 5–15% of patients develop severe sequelae called hemolytic uremic syndrome, which can be fatal [116].

The current state of knowledge reveals only a glimpse of the potential role that Gb3/CD77 synthase and its products play in the metastasis of cancer cells and their chemoresistance. Both TM9SF2 and GOLPH3, which are crucial for Gb3/CD77 synthase localization in the Golgi and hence influence its activity, were identified as oncoproteins. TM9SF2 has been implicated in the progression of colorectal and pancreatic cancer [117, 118]. GOLPH3 contributes to the tumorigenesis of lung, breast, colorectal, prostate, ovarian, gastric, and hepatocellular cancer, as well as melanoma and glioma, through controlling Golgi-to-plasma membrane transport and DNA damage response [119]. We hypothesize that GSL products of Gb3/CD77 synthase play significant roles in oncogenic axes mediated by GOLPH3 and TM9SF2, potentially opening up a new chapter in the complex and intriguing story of this glycosyltransferase.

Availability of data and materials

Not applicable.

Abbreviations

CD77:

Cluster of differentiation 77

CRISPR:

Clustered regularly interspaced short palindromic repeats

CTH:

Ceramide trihexoside

EGR1:

Early growth response protein

EMT:

Epithelial-to-mesenchymal transition

Ga2:

Galabiosylceramide

Gb3:

Globotriaosylceramide

Gb4:

Globotetraosylceramide

GlcCer:

Glucosylceramide

GOLPH3:

Golgi phosphoprotein3

GSL:

Glycosphingolipid

ISBT:

International Society of Blood Transfusion

LAPTM4A:

Lysosome-associated protein transmembrane 4α

mRNA:

Messenger RNA

ORF:

Open reading frame

RBC:

Red blood cel

RUNX1:

Runt-related transcription factor1

SEB:

Staphylococcal enterotoxin B

siRNA:

Small interfering RNA

SNFG:

Symbol Nomenclature for Glycans

SNP:

Single-nucleotide polymorphism

STEC:

Shiga toxin-producing Escherichia coli

TGN:

Trans-Golgi network

TM9SF2:

Transmembrane 9 superfamily 2 protein

UPEC:

Uropathogenic Escherichia coli

References

  1. Albesa-Jové D, Guerin ME. The conformational plasticity of glycosyltransferases. Curr Opin Struct Biol. 2016;40:23–32. https://doi.org/10.1016/j.sbi.2016.07.007.

    Article  CAS  PubMed  Google Scholar 

  2. Jacob F, Alam S, Konantz M, Liang C, Kohler RS, Everest-Dass AV, et al. Transition of mesenchymal and epithelial cancer cells depends on α1-4 galactosyltransferase-mediated glycosphingolipids. Cancer Res. 2018;78:2952–65. https://doi.org/10.1158/0008-5472.CAN-17-2223.

    Article  CAS  PubMed  Google Scholar 

  3. Mikolajczyk K, Bereznicka A, Szymczak-Kulus K, Haczkiewicz-Lesniak K, Szulc B, Olczak M, et al. Missing the sweet spot: one of the two N-glycans on human Gb3/CD77 synthase is expendable. Glycobiology. 2021;31:1145–62. https://doi.org/10.1093/glycob/cwab041.

    Article  CAS  PubMed  Google Scholar 

  4. Mikolajczyk K, Sikora M, Hanus C, Kaczmarek R, Czerwinski M. One of the two N-glycans on the human Gb3/CD77 synthase is essential for its activity and allosterically regulates its function. Biochem Biophys Res Commun. 2022;617:36–41. https://doi.org/10.1016/j.bbrc.2022.05.085.

    Article  CAS  PubMed  Google Scholar 

  5. Yamaji T, Sekizuka T, Tachida Y, Sakuma C, Morimoto K, Kuroda M, et al. A CRISPR screen identifies LAPTM4A and TM9SF proteins as glycolipid-regulating factors. IScience. 2019;11:409–24. https://doi.org/10.1016/j.isci.2018.12.039.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. D’Angelo G, Uemura T, Chuang CC, Polishchuk E, Santoro M, Ohvo-Rekilä H, et al. Vesicular and non-vesicular transport feed distinct glycosylation pathways in the Golgi. Nature. 2013;501:116–20. https://doi.org/10.1038/nature12423.

    Article  CAS  PubMed  Google Scholar 

  7. Rizzo R, Russo D, Kurokawa K, Sahu P, Lombardi B, Supino D, et al. Golgi maturation-dependent glycoenzyme recycling controls glycosphingolipid biosynthesis and cell growth via GOLPH3. EMBO J. 2021;40:1–21. https://doi.org/10.15252/embj.2020107238.

    Article  CAS  Google Scholar 

  8. Tian S, Muneeruddin K, Choi MY, Tao L, Bhuiyan RH, Ohmi Y, et al. Genome-wide CRISPR screens for Shiga toxins and ricin reveal Golgi proteins critical for glycosylation. PLOS Biol. 2018;16: e2006951. https://doi.org/10.1371/journal.pbio.2006951.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Welch LG, Munro S. A tale of short tails, through thick and thin: investigating the sorting mechanisms of Golgi enzymes. FEBS Lett. 2019;593:2452–65. https://doi.org/10.1002/1873-3468.13553.

    Article  CAS  PubMed  Google Scholar 

  10. Welch LG, Peak-Chew S-Y, Begum F, Stevens TJ, Munro S. GOLPH3 and GOLPH3L are broad-spectrum COPI adaptors for sorting into intra-Golgi transport vesicles. J Cell Biol. 2021. https://doi.org/10.1083/jcb.202106115.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Kojima Y, Fukumoto S, Furukawa K, Okajima T, Wiels J, Yokoyama K, et al. Molecular cloning of globotriaosylceramide/CD77 synthase, a glycosyltransferase that initiates the synthesis of globo series glycosphingolipids. J Biol Chem. 2000;275:15152–6.

    Article  CAS  PubMed  Google Scholar 

  12. Kaczmarek R, Duk M, Szymczak K, Korchagina E, Tyborowska J, Mikolajczyk K, et al. Human Gb3/CD77 synthase reveals specificity toward two or four different acceptors depending on amino acid at position 211, creating Pk, P1 and NOR blood group antigens. Biochem Biophys Res Commun. 2016;470:168–74. https://doi.org/10.1016/j.bbrc.2016.01.017.

    Article  CAS  PubMed  Google Scholar 

  13. Szymczak-Kulus K, Weidler S, Bereznicka A, Mikolajczyk K, Kaczmarek R, Bednarz B, et al. Human Gb3/CD77 synthase produces P1 glycotope-capped N-glycans, which mediate Shiga toxin 1 but not Shiga toxin 2 cell entry. J Biol Chem. 2021;296:100299. https://doi.org/10.1016/j.jbc.2021.100299.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Jin C, Teneberg S. Characterization of novel nonacid glycosphingolipids as biomarkers of human gastric adenocarcinoma. J Biol Chem. 2022;298:101732. https://doi.org/10.1016/j.jbc.2022.101732.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Suchanowska A, Kaczmarek R, Duk M, Lukasiewicz J, Smolarek D, Majorczyk E, et al. A single point mutation in the gene encoding Gb3/CD77 synthase causes a rare inherited polyagglutination syndrome. J Biol Chem. 2012;287:38220–30. https://doi.org/10.1074/jbc.M112.408286.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Duk M, Reinhold BB, Reinhold VN, Kusnierz-Alejska G, Lisowska E. Structure of a neutral glycosphingolipid recognized by human antibodies in polyagglutinable erythrocytes from the rare NOR phenotype. J Biol Chem. 2001;276:40574–82. https://doi.org/10.1074/jbc.M102711200.

    Article  CAS  PubMed  Google Scholar 

  17. Czerwinski M, Kaczmarek R. NOR blood group antigen. In: Polyagglutination: an update and review. Bethesda: AABB Press; 2022. p. 63–72.

    Google Scholar 

  18. Harris PA, Roman GK, Moulds JJ, Bird GWG, Shah NG. An inherited RBC characteristic, NOR, resulting in erythrocyte polyagglutination. Vox Sang. 1982;42:134–40. https://doi.org/10.1111/j.1423-0410.1982.tb01083.x.

    Article  CAS  PubMed  Google Scholar 

  19. Kuśnierz-Alejska G, Duk M, Stony JR, Reid ME, Wiȩcek B, Seyfried H, et al. NOR polyagglutination and Sta glycophorin in one family: relation of NOR polyagglutination to terminal α-galactose residues and abnormal glycolipids. Transfusion. 1999;39:32–8. https://doi.org/10.1046/j.1537-2995.1999.39199116892.x.

    Article  PubMed  Google Scholar 

  20. Lisowska E, Duk M. Polyagglutination NOR: new glycosphingolipid antigens recognized by a new type of common human anti-α-galactosyl antibodies. Arch Biochem Biophys. 2004;426:142–7. https://doi.org/10.1016/j.abb.2004.02.035.

    Article  CAS  PubMed  Google Scholar 

  21. Mourad R, Morelle W, Neveu A, Strecker G. Diversity of O-linked glycosylation patterns between species: characterization of 25 carbohydrate chains from oviducal mucins of Rana ridibunda. Eur J Biochem. 2001;268:1990–2003. https://doi.org/10.1046/j.1432-1327.2001.02071.x.

    Article  CAS  PubMed  Google Scholar 

  22. Fontaine T, Delangle A, Simenel C, Coddeville B, van Vliet SJ, van Kooyk Y, et al. Galactosaminogalactan, a new immunosuppressive polysaccharide of Aspergillus fumigatus. PLoS Pathog. 2011;7: e1002372. https://doi.org/10.1371/journal.ppat.1002372.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Hellberg Å. P1PK: a blood group system with an identity crisis. ISBT Sci Ser. 2020;15:40–5. https://doi.org/10.1111/voxs.12505.

    Article  Google Scholar 

  24. Kaczmarek R, Buczkowska A, Mikołajewicz K, Krotkiewski H, Czerwinski M. P1PK, GLOB, and FORS blood group systems and GLOB collection: biochemical and clinical aspects: do we understand it all yet? Transfus Med Rev. 2014;28:126–36. https://doi.org/10.1016/j.tmrv.2014.04.007.

    Article  PubMed  Google Scholar 

  25. Reid ME, Lomas-Francis C, Olsson ML. The blood group antigen factsbook. 3rd ed. Amsterdam: Elsevier Ltd.; 2012. https://doi.org/10.1016/C2011-0-69689-9.

    Book  Google Scholar 

  26. Hellberg Å, Steffensen R, Yahalom V, Sojka BN, Heier HE, Levene C, et al. Additional molecular bases of the clinically important p blood group phenotype. Transfusion. 2003;43:899–907. https://doi.org/10.1046/j.1537-2995.2003.00425.x.

    Article  CAS  PubMed  Google Scholar 

  27. Steffensen R, Carlier K, Wiels J, Levery SB, Stroud M, Cedergren B, et al. Cloning and expression of the Histo-blood group PkUDP-galactose: Galβ1–4Glcβ1-Cer α1,4-galactosyltransferase. J Biol Chem. 2000;275:16723–9. https://doi.org/10.1074/jbc.M000728200.

    Article  CAS  PubMed  Google Scholar 

  28. Iwamura K, Furukawa K, Uchikawa M, Sojka BN, Kojima Y, Wiels J, et al. The blood group P1 synthase gene is identical to the Gb3/CD77 synthase gene. J Biol Chem. 2003;278:44429–38. https://doi.org/10.1074/jbc.M301609200.

    Article  CAS  PubMed  Google Scholar 

  29. Thuresson B, Westman JS, Olsson ML. Identification of a novel A4GALT exon reveals the genetic basis of the P1/P2 histo-blood groups. Blood. 2011;117:678–87. https://doi.org/10.1182/blood-2010-08-301333.

    Article  CAS  PubMed  Google Scholar 

  30. Lai Y-J, Wu W-Y, Yang C-M, Yang L-R, Chu C-C, Chan Y-S, et al. A systematic study of single-nucleotide polymorphisms in the A4GALT gene suggests a molecular genetic basis for the P1/P2 blood groups. Transfusion. 2014;54:3222–31. https://doi.org/10.1111/trf.12771.

    Article  CAS  PubMed  Google Scholar 

  31. Kaczmarek R, Szymczak-Kulus K, Bereźnicka A, Mikołajczyk K, Duk M, Majorczyk E, et al. Single nucleotide polymorphisms in A4GALT spur extra products of the human Gb3/CD77 synthase and underlie the P1PK blood group system. PLoS ONE. 2018;13: e0196627. https://doi.org/10.1371/journal.pone.0196627.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Thinley J, Nathalang O, Chidtrakoon S, Intharanut K. Blood group P1 prediction using multiplex PCR genotyping of A4GALT among Thai blood donors. Transfus Med. 2021;31:48–54. https://doi.org/10.1111/tme.12749.

    Article  PubMed  Google Scholar 

  33. Yeh C-C, Chang C-J, Twu Y-C, Hung S-T, Tsai Y-J, Liao J-C, et al. The differential expression of the blood group P1–A4GALT and P2–A4GALT alleles is stimulated by the transcription factor early growth response 1. Transfusion. 2018;58:1054–64. https://doi.org/10.1111/trf.14515.

    Article  CAS  PubMed  Google Scholar 

  34. Westman JS, Stenfelt L, Vidovic K, Möller M, Hellberg Å, Kjellström S, et al. Allele-selective RUNX1 binding regulates P1 blood group status by transcriptional control of A4GALT. Blood. 2018. https://doi.org/10.1182/blood-2017-08-803080.

    Article  PubMed  Google Scholar 

  35. Poole J, Daniels G. Blood group antibodies and their significance in transfusion medicine. Transfus Med Rev. 2007;21:58–71. https://doi.org/10.1016/j.tmrv.2006.08.003.

    Article  PubMed  Google Scholar 

  36. Duk M, Kusnierz-Alejska G, Korchagina EY, Bovin NV, Bochenek S, Lisowska E. Anti-α-galactosyl antibodies recognizing epitopes terminating with α1,4-linked galactose: human natural and mouse monoclonal anti-NOR and anti-P1 antibodies. Glycobiology. 2005;15:109–18. https://doi.org/10.1093/oxfordjournals.glycob.a034964.

    Article  CAS  PubMed  Google Scholar 

  37. Di Ciaccio P, Cutts B, Alahakoon TI, Dennington PM, Soo LA, Curnow J. Clinical consequences of the extremely rare anti-PP1Pk isoantibodies in pregnancy: a case series and review of the literature. Vox Sang. 2021;116:591–600. https://doi.org/10.1111/vox.13042.

    Article  CAS  PubMed  Google Scholar 

  38. Volynsky P, Efremov R, Mikhalev I, Dobrochaeva K, Tuzikov A, Korchagina E, et al. Why human anti-Galα1–4Galβ1–4Glc natural antibodies do not recognize the trisaccharide on erythrocyte membrane? Molecular dynamics and immunochemical investigation. Mol Immunol. 2017;90:87–97. https://doi.org/10.1016/j.molimm.2017.06.247.

    Article  CAS  PubMed  Google Scholar 

  39. Huflejt ME, Vuskovic M, Vasiliu D, Xu H, Obukhova P, Shilova N, et al. Anti-carbohydrate antibodies of normal sera: findings, surprises and challenges. Mol Immunol. 2009;46:3037–49. https://doi.org/10.1016/j.molimm.2009.06.010.

    Article  CAS  PubMed  Google Scholar 

  40. Hozumi I, Nishizawa M, Ariga T, Miyatake T. Biochemical and clinical analysis of accumulated glycolipids in symptomatic heterozygotes of angiokeratoma corporis diffusum (Fabry’s disease) in comparison with hemizygotes. J Lipid Res. 1990;31:335–40. https://doi.org/10.1016/s0022-2275(20)43219-5.

    Article  CAS  PubMed  Google Scholar 

  41. Boutin M, Menkovic I, Martineau T, Vaillancourt-Lavigueur V, Toupin A, Auray-Blais C. Separation and analysis of lactosylceramide, galabiosylceramide, and globotriaosylceramide by LC-MS/MS in urine of Fabry disease patients. Anal Chem. 2017;89:13382–90. https://doi.org/10.1021/acs.analchem.7b03609.

    Article  CAS  PubMed  Google Scholar 

  42. Akiyama H, Ide M, Yamaji T, Mizutani Y, Niimi Y, Mutoh T, et al. Galabiosylceramide is present in human cerebrospinal fluid. Biochem Biophys Res Commun. 2021;536:73–9. https://doi.org/10.1016/j.bbrc.2020.12.051.

    Article  CAS  PubMed  Google Scholar 

  43. Cooling LL, Walker KE, Gille T, Koerner TA. Shiga toxin binds human platelets via globotriaosylceramide (Pk antigen) and a novel platelet glycosphingolipid. Infect Immun. 1998;66:4355–66.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Stenfelt L, Westman JS, Hellberg Å, Olsson ML. The P1 histo-blood group antigen is present on human red blood cell glycoproteins. Transfusion. 2019;59:1108–17. https://doi.org/10.1111/trf.15115.

    Article  CAS  PubMed  Google Scholar 

  45. Cooling L. Blood groups in infection and host susceptibility. Clin Microbiol Rev. 2015;28:801–70. https://doi.org/10.1128/CMR.00109-14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Oosterwlik E, Kalisiak A, Wakka JC, Scheinberg DA, Old LJ. Monoclonal antibodies against Galα 1–4Galß 1–4Glc(Pk, CD77) produced with a synthetic glycoconjugate as immunogen: reactivity with carbohydrates, with fresh frozen human tissues and hematopoietic tumors. Int J Cancer. 1991;48:848–54. https://doi.org/10.1002/ijc.2910480610.

    Article  Google Scholar 

  47. Zumbrun SD, Hanson L, Sinclair JF, Freedy J, Melton-Celsa AR, Rodriguez-Canales J, et al. Human intestinal tissue and cultured colonic cells contain globotriaosylceramide synthase mRNA and the alternate Shiga toxin receptor globotetraosylceramide. Infect Immun. 2010;78:4488–99. https://doi.org/10.1128/IAI.00620-10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Obata F, Tohyama K, Bonev AD, Kolling GL, Tiffany R, Gross LK, et al. Shiga toxin 2 affects the central nervous system through receptor Gb3 localized to neurons fumiko. J Infect Dis. 2008;198:1398–406. https://doi.org/10.1086/591911.Shiga.

    Article  CAS  PubMed  Google Scholar 

  49. Murray LJ, Habeshaw JA, Wiels J, Greaves MF. Expression of Burkitt lymphoma-associated antigen (defined by the monoclonal antibody 38.13) on both normal and malignant germinal-centre B cells. Int J Cancer. 1985;36:561–5. https://doi.org/10.1002/ijc.2910360508.

    Article  CAS  PubMed  Google Scholar 

  50. Schwartz-Albiez R, Dörken B, Möller P, Brodin NT, Monner DA, Kniep B. Neutral glycosphingolipids of the globo-series characterize activation stages corresponding to germinal center B cells. Int Immunol. 1990;2:929–36. https://doi.org/10.1093/intimm/2.10.929.

    Article  CAS  PubMed  Google Scholar 

  51. Mangeney M, Richard Y, Coulaud D, Tursz T, Wiels J. CD77: an antigen of germinal center B cells entering apoptosis. Eur J Immunol. 1991;21:1131–40. https://doi.org/10.1002/eji.1830210507.

    Article  CAS  PubMed  Google Scholar 

  52. Taga S, Carlier K, Mishal Z, Capoulade C, Mangeney M, Lécluse Y, et al. Intracellular signaling events in CD77-mediated apoptosis of Burkitt’s lymphoma cells. Blood. 1997;90:2757–67. https://doi.org/10.1182/blood.v90.7.2757.

    Article  CAS  PubMed  Google Scholar 

  53. Sharma P, Zhang X, Ly K, Zhang Y, Hu Y, Ye AY, et al. The lipid globotriaosylceramide promotes germinal center B cell responses and antiviral immunity. Science. 2024;383:720–34. https://doi.org/10.1126/science.adg0564.

    Article  CAS  Google Scholar 

  54. Zhang T, De Waard AA, Wuhrer M, Spaapen RM. The role of glycosphingolipids in immune cell functions. Front Immunol. 2019;10:1–22. https://doi.org/10.3389/fimmu.2019.00090.

    Article  CAS  Google Scholar 

  55. Porubsky S, Federico G, Müthing J, Jennemann R, Gretz N, Büttner S, et al. Direct acute tubular damage contributes to Shigatoxin-mediated kidney failure. J Pathol. 2014;234:120–33. https://doi.org/10.1002/path.4388.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Morace I, Pilz R, Federico G, Jennemann R, Krunic D, Nordström V, et al. Renal globotriaosylceramide facilitates tubular albumin absorption and its inhibition protects against acute kidney injury. Kidney Int. 2019;96:327–41. https://doi.org/10.1016/j.kint.2019.02.010.

    Article  CAS  PubMed  Google Scholar 

  57. Newburg DS, Ashkenazi S, Cleary TG. Human milk contains the shiga toxin and Shiga-like toxin receptor glycolipid Gb3. J Infect Dis. 1992;166:832–6. https://doi.org/10.1093/infdis/166.4.832.

    Article  CAS  PubMed  Google Scholar 

  58. Dotz V, Wuhrer M. Histo-blood group glycans in the context of personalized medicine. Biochim Biophys Acta Gen Subj. 2016;1860:1596–607. https://doi.org/10.1016/j.bbagen.2015.12.026.

    Article  CAS  Google Scholar 

  59. Kok K, Zwiers KC, Boot RG, Overkleeft HS, Aerts JMFG, Artola M. Fabry disease: molecular basis, pathophysiology, diagnostics and potential therapeutic directions. Biomolecules. 2021;11:271. https://doi.org/10.3390/biom11020271.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Van Eijk M, Ferra MJ, Boot RG, Aerts JMFG. Lyso-glycosphingolipids: presence and consequences. Essays Biochem. 2020;64:565–78. https://doi.org/10.1042/EBC20190090.

    Article  PubMed  PubMed Central  Google Scholar 

  61. Miller JJ, Kanack AJ, Dahms NM. Progress in the understanding and treatment of Fabry disease. Biochim Biophys Acta Gen Subj. 2020;1864:129437. https://doi.org/10.1016/j.bbagen.2019.129437.

    Article  CAS  PubMed  Google Scholar 

  62. Yogasundaram H, Kim D, Oudit O, Thompson RB, Weidemann F, Oudit GY. Clinical features, diagnosis, and management of patients with Anderson-Fabry cardiomyopathy. Can J Cardiol. 2017;33:883–97. https://doi.org/10.1016/j.cjca.2017.04.015.

    Article  PubMed  Google Scholar 

  63. Germain DP. Fabry disease. Orphanet J Rare Dis. 2010;5:30. https://doi.org/10.1186/1750-1172-5-30.

    Article  PubMed  PubMed Central  Google Scholar 

  64. Ishii S, Taguchi A, Okino N, Ito M, Maruyama H. Determination of globotriaosylceramide analogs in the organs of a mouse model of Fabry disease. J Biol Chem. 2020;295:5577–87. https://doi.org/10.1074/jbc.RA120.012665.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Mehta A, Clarke JTR, Giugliani R, Elliott P, Linhart A, Beck M, et al. Natural course of Fabry disease: changing pattern of causes of death in FOS—Fabry Outcome Survey. J Med Genet. 2009;46:548–52. https://doi.org/10.1136/jmg.2008.065904.

    Article  CAS  PubMed  Google Scholar 

  66. Palaiodimou L, Kokotis P, Zompola C, Papagiannopoulou G, Bakola E, Papadopoulou M, et al. Fabry disease: current and novel therapeutic strategies: a narrative review. Curr Neuropharmacol. 2023;21:440–56. https://doi.org/10.2174/1570159X20666220601124117.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Cui S, Shin YJ, Fang X, Lee H, Eum SH, Ko EJ, et al. CRISPR/Cas9-mediated A4GALT suppression rescues Fabry disease phenotypes in a kidney organoid model. Transl Res. 2023;258:35–46. https://doi.org/10.1016/j.trsl.2023.02.005.

    Article  CAS  PubMed  Google Scholar 

  68. Shin YJ, Chae SY, Lee H, Fang X, Cui S, Lim SW, et al. CRISPR/Cas9-mediated suppression of A4GALT rescues endothelial cell dysfunction in a Fabry disease vasculopathy model derived from human induced pluripotent stem cells. Atherosclerosis. 2024;397:118549. https://doi.org/10.1016/j.atherosclerosis.2024.118549.

    Article  CAS  PubMed  Google Scholar 

  69. Siukstaite L, Imberty A, Römer W. Structural diversities of lectins binding to the glycosphingolipid Gb3. Front Mol Biosci. 2021;8:1–16. https://doi.org/10.3389/fmolb.2021.704685.

    Article  CAS  Google Scholar 

  70. Bereznicka A, Mikolajczyk K, Czerwinski M, Kaczmarek R. Microbial lectome versus host glycolipidome: how pathogens exploit glycosphingolipids to invade, dupe or kill. Front Microbiol. 2022;13:1–21. https://doi.org/10.3389/fmicb.2022.958653.

    Article  Google Scholar 

  71. Johannes L, Römer W. Shiga toxins—from cell biology to biomedical applications. Nat Rev Microbiol. 2010;8:105–16. https://doi.org/10.1038/nrmicro2279.

    Article  CAS  PubMed  Google Scholar 

  72. Beddoe T, Paton AW, Le Nours J, Rossjohn J, Paton JC. Structure, biological functions and applications of the AB5 toxins. Trends Biochem Sci. 2010;35:411–8. https://doi.org/10.1016/j.tibs.2010.02.003.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Cody EM, Dixon BP. Hemolytic uremic syndrome. Pediatr Clin North Am. 2019;66:235–46. https://doi.org/10.1016/j.pcl.2018.09.011.

    Article  PubMed  Google Scholar 

  74. Morimoto K, Suzuki N, Tanida I, Kakuta S, Furuta Y, Uchiyama Y, et al. Blood group P1 antigen–bearing glycoproteins are functional but less efficient receptors of Shiga toxin than conventional glycolipid-based receptors. J Biol Chem. 2020;295:9490–501. https://doi.org/10.1074/jbc.RA120.013926.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. Lindberg AA, Brown JE, Strömberg N, Westling-Ryd M, Schultz JE, Karlsson KA. Identification of the carbohydrate receptor for Shiga toxin produced by Shigella dysenteriae type 1. J Biol Chem. 1987;262:1779–85.

    Article  CAS  PubMed  Google Scholar 

  76. Brown JE, Echeverria P, Lindberg AA. Digalactosyl-containing glycolipids as cell surface receptors for Shiga toxin of Shigella dysenteriae 1 and related cytotoxins of Escherichia coli. Clin Infect Dis. 1991;13:S298-303. https://doi.org/10.1093/clinids/13.Supplement_4.S298.

    Article  CAS  Google Scholar 

  77. Szymczak-Kulus K. Human α-1,4-galactosyltransferase as an enzyme regulating Shiga toxin binding. [doctoral dissertation] Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences. 2021. https://bip.iitd.pan.wroc.pl/system/obj/1036_Katarzyna_Szymczak_doktorat.pdf. [Accessed October 3, 2024]

  78. Chatterjee S, Khullar M, Shi WY. Digalactosylceramide is the receptor for staphylococcal enterotoxin-B in human kidney proximal tubular cells. Glycobiology. 1995;5:327–33. https://doi.org/10.1093/glycob/5.3.327.

    Article  CAS  PubMed  Google Scholar 

  79. Legros N, Ptascheck S, Pohlentz G, Karch H, Dobrindt U, Müthing J. PapG subtype-specific binding characteristics of Escherichia coli towards globo-series glycosphingolipids of human kidney and bladder uroepithelial cells. Glycobiology. 2019;29:789–802. https://doi.org/10.1093/glycob/cwz059.

    Article  CAS  PubMed  Google Scholar 

  80. Bock K, Breimer ME, Brignole A, Hansson GC, Karlsson KA, Larson G, et al. Specificity of binding of a strain of uropathogenic Escherichia coli to Gal alpha 1—-4Gal-containing glycosphingolipids. J Biol Chem. 1985;260:8545–51. https://doi.org/10.1016/S0021-9258(17)39507-8.

    Article  CAS  PubMed  Google Scholar 

  81. Kouki A, Haataja S, Loimaranta V, Pulliainen AT, Nilsson UJ, Finne J. Identification of a novel streptococcal adhesin P (SadP) protein recognizing galactosyl-α1–4-galactose-containing glycoconjugates. J Biol Chem. 2011;286:38854–64. https://doi.org/10.1074/jbc.M111.260992.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  82. Lanne B, Cîopraga J, Bergström J, Motas C, Karlsson KA. Binding of the galactose-specific Pseudomonas aeruginosa lectin, PA-I, to glycosphingolipids and other glycoconjugates. Glycoconj J. 1994;11:292–8. https://doi.org/10.1007/BF00731201.

    Article  CAS  PubMed  Google Scholar 

  83. Blanchard B, Nurisso A, Hollville E, Tétaud C, Wiels J, Pokorná M, et al. Structural basis of the preferential binding for globo-series glycosphingolipids displayed by Pseudomonas aeruginosa lectin I. J Mol Biol. 2008;383:837–53. https://doi.org/10.1016/j.jmb.2008.08.028.

    Article  CAS  PubMed  Google Scholar 

  84. Beshr G, Sikandar A, Jemiller E-M, Klymiuk N, Hauck D, Wagner S, et al. Photorhabdus luminescens lectin A (PllA): a new probe for detecting α-galactoside–terminating glycoconjugates. J Biol Chem. 2017;292:19935–51. https://doi.org/10.1074/jbc.M117.812792.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Nudelman E, Kannagi R, Hakomori S, Parsons M, Lipinski M, Wiels J, et al. A glycolipid antigen associated with Burkitt lymphoma defined by a monoclonal antibody. Science. 1983;220:509–11. https://doi.org/10.1126/science.6836295.

    Article  CAS  PubMed  Google Scholar 

  86. Wiels J, Fellous M, Tursz T. Monoclonal antibody against a Burkitt lymphoma-associated antigen. Proc Natl Acad Sci USA. 1981;78:6485–8. https://doi.org/10.1073/pnas.78.10.6485.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  87. Kovbasnjuk O, Mourtazina R, Baibakov B, Wang T, Elowsky C, Choti MA, et al. The glycosphingolipid globotriaosylceramide in the metastatic transformation of colon cancer. Proc Natl Acad Sci. 2005;102:19087–92. https://doi.org/10.1073/pnas.0506474102.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Falguieres T, Maak M, von Weyhern C, Sarr M, Sastre X, Poupon M-F, et al. Human colorectal tumors and metastases express Gb3 and can be targeted by an intestinal pathogen-based delivery tool. Mol Cancer Ther. 2008;7:2498–508. https://doi.org/10.1158/1535-7163.MCT-08-0430.

    Article  CAS  PubMed  Google Scholar 

  89. Distler U, Souady J, Hülsewig M, Drmić-Hofman I, Haier J, Friedrich AW, et al. Shiga toxin receptor Gb3Cer/CD77: tumor-association and promising therapeutic target in pancreas and colon cancer. PLoS ONE. 2009;4: e6813. https://doi.org/10.1371/journal.pone.0006813.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  90. Geyer PE, Maak M, Nitsche U, Perl M, Novotny A, Slotta-Huspenina J, et al. Gastric adenocarcinomas express the glycosphingolipid Gb3/CD77: targeting of gastric cancer cells with Shiga toxin B-subunit. Mol Cancer Ther. 2016;15:1008–17. https://doi.org/10.1158/1535-7163.MCT-15-0633.

    Article  CAS  PubMed  Google Scholar 

  91. Sawada R, Hotta H, Chung YS, Sowa M, Tai T, Yano I. Globotriaosyl ceramide and globoside as major glycolipid components of fibroblasts in scirrhous gastric carcinoma tissues. Japanese J Cancer Res. 1998;89:167–76. https://doi.org/10.1111/j.1349-7006.1998.tb00545.x.

    Article  CAS  Google Scholar 

  92. Maak M, Nitsche U, Keller L, Wolf P, Sarr M, Thiebaud M, et al. Tumor-specific targeting of pancreatic cancer with shiga toxin B-subunit. Mol Cancer Ther. 2011;10:1918–28. https://doi.org/10.1158/1535-7163.MCT-11-0006.

    Article  CAS  PubMed  Google Scholar 

  93. LaCasse EC, Bray MR, Patterson B, Lim WM, Perampalam S, Radvanyi LG, et al. Shiga-like toxin-1 receptor on human breast cancer, lymphoma, and myeloma and absence from CD34+ hematopoietic stem cells: implications for ex vivo tumor purging and autologous stem cell transplantation. Blood. 1999;94:2901–10.

    CAS  PubMed  Google Scholar 

  94. Johansson D, Kosovac E, Moharer J, Ljuslinder I, Brännström T, Johansson A, et al. Expression of verotoxin-1 receptor Gb3 in breast cancer tissue and verotoxin-1 signal transduction to apoptosis. BMC Cancer. 2009;9:67. https://doi.org/10.1186/1471-2407-9-67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  95. Stimmer L, Dehay S, Nemati F, Massonnet G, Richon S, Decaudin D, et al. Human breast cancer and lymph node metastases express Gb3 and can be targeted by STxB-vectorized chemotherapeutic compounds. BMC Cancer. 2014;14:916.

    Article  PubMed  PubMed Central  Google Scholar 

  96. Arab S, Russel E, Chapman WB, Rosen B, Lingwood CA. Expression of the verotoxin receptor glycolipid, globotriaosylceramide, in ovarian hyperplasias. Oncol Res. 1997;9:553–63.

    CAS  PubMed  Google Scholar 

  97. Cumin C, Huang Y, Rossdam C, Ruoff F, Céspedes SP, Liang C-Y, et al. Glycosphingolipids are mediators of cancer plasticity through independent signaling pathways. Cell Rep. 2022;40:111181. https://doi.org/10.1016/j.celrep.2022.111181.

    Article  CAS  PubMed  Google Scholar 

  98. Robert A, Wiels J. Shiga toxins as antitumor tools. Toxins. 2021;13:1–15. https://doi.org/10.3390/toxins13100690.

    Article  CAS  Google Scholar 

  99. Jacob F, Anugraham M, Pochechueva T, Tse BWC, Alam S, Guertler R, et al. The glycosphingolipid P1 is an ovarian cancer-associated carbohydrate antigen involved in migration. Br J Cancer. 2014;111:1634–45. https://doi.org/10.1038/bjc.2014.455.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Hořejší K, Jin C, Vaňková Z, Jirásko R, Strouhal O, Melichar B, et al. Comprehensive characterization of complex glycosphingolipids in human pancreatic cancer tissues. J Biol Chem. 2023. https://doi.org/10.1016/j.jbc.2023.102923.

    Article  PubMed  PubMed Central  Google Scholar 

  101. Cumin C, Huang YL, Everest-Dass A, Jacob F. Deciphering the importance of glycosphingolipids on cellular and molecular mechanisms associated with epithelial-to-mesenchymal transition in cancer. Biomolecules. 2021;11:1–19. https://doi.org/10.3390/biom11010062.

    Article  CAS  Google Scholar 

  102. Zhang T, van Die I, Tefsen B, van Vliet SJ, Laan LC, Zhang J, et al. Differential O- and glycosphingolipid glycosylation in human pancreatic adenocarcinoma cells with opposite morphology and metastatic behavior. Front Oncol. 2020;10:1–19. https://doi.org/10.3389/fonc.2020.00732.

    Article  Google Scholar 

  103. Liu Y-Y, Hill RA, Li Y-T. Ceramide glycosylation catalyzed by glucosylceramide synthase and cancer drug resistance. Adv Cancer Res. 2013;117:59–89. https://doi.org/10.1016/B978-0-12-394274-6.00003-0.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  104. Liu Y-Y, Gupta V, Patwardhan GA, Bhinge K, Zhao Y, Bao J, et al. Glucosylceramide synthase upregulates MDR1 expression in the regulation of cancer drug resistance through cSrc and β-catenin signaling. Mol Cancer. 2010;9:1–14. https://doi.org/10.1186/1476-4598-9-145.

    Article  CAS  Google Scholar 

  105. Wegner MS, Schömel N, Gruber L, Örtel SB, Kjellberg MA, Mattjus P, et al. UDP-glucose ceramide glucosyltransferase activates AKT, promoted proliferation, and doxorubicin resistance in breast cancer cells. Cell Mol Life Sci. 2018;75:3393–410. https://doi.org/10.1007/s00018-018-2799-7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  106. Roy KR, Uddin MB, Roy SC, Hill RA, Marshall J, Li YT, et al. Gb3-cSrc complex in glycosphingolipid-enriched microdomains contributes to the expression of p53 mutant protein and cancer drug resistance via β-catenin–activated RNA methylation. FASEB BioAdv. 2020;2:653–67. https://doi.org/10.1096/fba.2020-00044.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  107. Schwab M, Multhoff G. A low membrane Hsp70 expression in tumor cells with impaired lactate metabolism mediates radiosensitization by NVP-AUY922. Front Oncol. 2022;12:1–7. https://doi.org/10.3389/fonc.2022.861266.

    Article  CAS  Google Scholar 

  108. Multhoff G, Pockley AG, Schmid TE, Schilling D. The role of heat shock protein 70 (Hsp70) in radiation-induced immunomodulation. Cancer Lett. 2015;368:179–84. https://doi.org/10.1016/j.canlet.2015.02.013.

    Article  CAS  PubMed  Google Scholar 

  109. Sugawara S, Kawano T, Omoto T, Hosono M, Tatsuta T, Nitta K. Binding of Silurus asotus lectin to Gb3 on Raji cells causes disappearance of membrane-bound form of HSP70. Biochim Biophys Acta Gen Subj. 2009;1790:101–9. https://doi.org/10.1016/j.bbagen.2008.10.005.

    Article  CAS  Google Scholar 

  110. Gehrmann M, Liebisch G, Schmitz G, Anderson R, Steinem C, De Maio A, et al. Tumor-specific Hsp70 plasma membrane localization is enabled by the glycosphingolipid Gb3. PLoS ONE. 2008;3:1–9. https://doi.org/10.1371/journal.pone.0001925.

    Article  CAS  Google Scholar 

  111. Watkins WM, Greenwell P, Yates AD, Johnson PH. Regulation of expression of carbohydrate blood group antigens. Biochimie. 1988;70:1597–611. https://doi.org/10.1016/0300-9084(88)90295-7.

    Article  CAS  PubMed  Google Scholar 

  112. Maccioni HJF, Quiroga R, Ferrari ML. Cellular and molecular biology of glycosphingolipid glycosylation. J Neurochem. 2011;117:589–602.

    CAS  PubMed  Google Scholar 

  113. Kellokumpu S, Hassinen A, Glumoff T. Glycosyltransferase complexes in eukaryotes: long-known, prevalent but still unrecognized. Cell Mol Life Sci. 2016;73:305–25. https://doi.org/10.1007/s00018-015-2066-0.

    Article  CAS  PubMed  Google Scholar 

  114. Mikołajczyk K, Wróblewski K, Kmiecik S. Delving into human α1,4-galactosyltransferase acceptor specificity: the role of enzyme dimerization. Biochem Biophys Res Commun. 2024;736:150486. https://doi.org/10.1016/j.bbrc.2024.150486.

    Article  CAS  PubMed  Google Scholar 

  115. Majowicz SE, Scallan E, Jones-Bitton A, Sargeant JM, Stapleton J, Angulo FJ, et al. Global incidence of human Shiga toxin-producing Escherichia coli infections and deaths: a systematic review and knowledge synthesis. Foodborne Pathog Dis. 2014;11:447–55. https://doi.org/10.1089/fpd.2013.1704.

    Article  PubMed  Google Scholar 

  116. Mayer CL, Leibowitz CS, Kurosawa S, Stearns-Kurosawa DJ. Shiga toxins and the pathophysiology of hemolytic uremic syndrome in humans and animals. Toxins. 2012;4:1261–87. https://doi.org/10.3390/toxins4111261.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  117. Clark CR, Maile M, Blaney P, Hellweg SR, Strauss A, Durose W, et al. Transposon mutagenesis screen in mice identifies TM9SF2 as a novel colorectal cancer oncogene. Sci Rep. 2018;8:1–12. https://doi.org/10.1038/s41598-018-33527-3.

    Article  CAS  Google Scholar 

  118. Li Q, Lei C, Lu C, Wang J, Gao M, Gao W. LINC01232 exerts oncogenic activities in pancreatic adenocarcinoma via regulation of TM9SF2. Cell Death Dis. 2019. https://doi.org/10.1038/s41419-019-1896-3.

    Article  PubMed  PubMed Central  Google Scholar 

  119. Sechi S, Frappaolo A, Karimpour-Ghahnavieh A, Piergentili R, Giansanti MG. Oncogenic roles of GOLPH3 in the physiopathology of cancer. Int J Mol Sci. 2020;21:933. https://doi.org/10.3390/ijms21030933.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  120. Neelamegham S, Aoki-Kinoshita K, Bolton E, Frank M, Lisacek F, Lütteke T, et al. Updates to the symbol nomenclature for glycans guidelines. Glycobiology. 2019;29:620–4. https://doi.org/10.1093/glycob/cwz045.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Łukasz Sobala for phylogenetic analysis of A4GALT genes.

Funding

This research received no external funding.

Author information

Authors and Affiliations

  1. Laboratory of Glycobiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114, Wroclaw, Poland

    Katarzyna Szymczak-Kulus, Marcin Czerwinski & Radoslaw Kaczmarek

Authors
  1. Katarzyna Szymczak-Kulus
    View author publications

    Search author on:PubMed Google Scholar

  2. Marcin Czerwinski
    View author publications

    Search author on:PubMed Google Scholar

  3. Radoslaw Kaczmarek
    View author publications

    Search author on:PubMed Google Scholar

Contributions

Conceptualization, all authors; writing—original draft preparation, K.S.-K.; writing—review and editing, all authors; visualization, K.S.-K. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Katarzyna Szymczak-Kulus.

Ethics declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Szymczak-Kulus, K., Czerwinski, M. & Kaczmarek, R. Human Gb3/CD77 synthase: a glycosyltransferase at the crossroads of immunohematology, toxicology, and cancer research. Cell Mol Biol Lett 29, 137 (2024). https://doi.org/10.1186/s11658-024-00658-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s11658-024-00658-7

Keywords

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us