Differential carbohydrate binding and cell surface glycosylation of human cancer cell lines

Arndt, Nadia X.; Tiralongo, Joe; Madge, Paul D.; Itzstein, Mark von; Day, Christopher J.

doi:10.1002/jcb.23139

Cited by 53 publications

(53 citation statements)

References 27 publications

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“…Glycan arrays were prepared as per Arndt et al (64). For whole bacteria, 10 6 cells were labeled with CFDA-SE, Bodipy 558-SE, or FITC for 30 min in 1× PBS with washing and application to the array performed as described by Day et al (52).…”

Section: Methodsmentioning

confidence: 99%

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Day

Tran

Semchenko

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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108

104

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Cells from all domains of life express glycan structures attached to lipids and proteins on their surface, called glycoconjugates. Cell-tocell contact mediated by glycan:glycan interactions have been considered to be low-affinity interactions that precede highaffinity protein-glycan or protein-protein interactions. In several pathogenic bacteria, truncation of surface glycans, lipooligosaccharide (LOS), or lipopolysaccharide (LPS) have been reported to significantly reduce bacterial adherence to host cells. Here, we show that the saccharide component of LOS/LPS have direct, high-affinity interactions with host glycans. Glycan microarrays reveal that LOS/LPS of four distinct bacterial pathogens bind to numerous host glycan structures. Surface plasmon resonance was used to determine the affinity of these interactions and revealed 66 high-affinity host-glycan:bacterial-glycan pairs with equilibrium dissociation constants (K D ) ranging between 100 nM and 50 μM. These glycan:glycan affinity values are similar to those reported for lectins or antibodies with glycans. Cell assays demonstrated that glycan:glycan interaction-mediated bacterial adherence could be competitively inhibited by either host cell or bacterial glycans. This is the first report to our knowledge of high affinity glycan:glycan interactions between bacterial pathogens and the host. The discovery of large numbers of glycan:glycan interactions between a diverse range of structures suggests that these interactions may be important in all biological systems.H ost surface glycosylation is ubiquitous and is targeted by pathogenic bacteria, viruses, fungi and parasites for adherence and toxin binding and by glycosidases (1). Escherichia coli type 1 fimbriae, FimH, is one of the most widely studied glycanrecognizing protein adhesins, with specificity for monomannose to oligomannose structures with the variability of the mannose structure bound leading to different tissue tropism (2). Other glycan-recognizing adhesins expressed by bacteria include the following: Pseudomonas aeruginosa lectins 1 and 2 (PA-IL and PA-IIL) that have specificity for galactose and fucose, respectively (3); Helicobacter pylori SabA, specific for sialic acid containing glycoconjugates including sialyLewis X; and BabAspecific for fucosylated glycoconjugates including Lewis B (4, 5). Although there are numerous known glycan binding adhesins, the adhesins of some bacteria that interact with host surface glycans remain unknown.Direct interactions between surface glycans (glycan:glycan interactions) have been reported in sea sponges as heterogenous glycan interactions, and in mouse embryo development and cancer where homodimers of Lewis X (LeX) or ganglioside structures play a role in cell adhesion and growth factor receptor interactions (6, 7). Outside of these reports, glycan:glycan interactions, when noted, have generally been considered to be low-affinity, weak interactions (8) that precede high-affinity protein:glycan or protein:protein interactions (1, 2, 5, 9).Interestingly, there...

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Section: Methodsmentioning

confidence: 99%

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Day

Tran

Semchenko

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

108

104

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“…Thus, the interactions of the C. jejuni proteins with various metabolites, including some involved in the TCA cycle, were investigated. To this end, a TCA array was developed based on a technique previously used to analyze interactions of various proteins with glycans, amino acids and lectins [29,31,40]. The TCA array was constructed and verified by hybridization with human glutathione reductase, a well characterized and commercially available protein.…”

Section: Discussionmentioning

confidence: 99%

Identification of NuoX and NuoY Ligand Binding Specificity in the Campylobacter Jejuni Complex I

Calderón-Gómez¹,

Day²,

Hartley-Tassell³

et al. 2017

J Bacteriol Parasitol

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The components of the proton pump NADH:ubiquinone (Complex I) of the respiration pathway have been identified in the C. jejuni genome. However, the paradigm genes nuoE and nuoF encoding subunits of the NADH dehydrogenase module of Complex I are absent. Instead the genes cj1575c and cj1574c encoding NuoX and NuoY are present in the loci corresponding to nuoE and nuoF, respectively. Bioinformatics analyses showed the presence of nuoX and nuoY homologues in all sequenced strains of C. jejuni and in other Campylobacter species, as well as the presence of orthologues in other ɛ-Proteobacteria.To understand the involvement of the NuoX and NuoY proteins in the respiration of C. jejuni and to characterize their ligand binding specificity and affinity, a tricarboxylic acid cycle array was developed as a tool to identify proteins that can bind to intermediates of this cycle as well as other metabolites. This array showed that NuoX bound FAD 2+ , and NuoY bound FAD 2+ and the electron donors malate and lactate. Saturation Transfer Difference Nuclear Magnetic Resonance studies confirmed the NuoY binding ligands, and suggested that the flavin moiety of FAD 2+ interacted more strongly with NuoY than the adenine moiety. Affinity data generated by Surface Plasmon Resonance indicated that NuoY bound to FAD 2+ with a K D of 337 nM; NuoX and NuoY had an affinity for NADH of a K D of 403 nM and 478 nM, respectively, and a ten-fold lower affinity for both NAD + and FAD 2+ . The data suggested that the flavin-adenine dinucletoide could be bound preferentially to the NAD in the Complex I of C. jejuni.

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“…Glycan arrays consisting of 367 diverse glycans with and without the presence of one of three spacers (sp2, sp3 or sp4 20,48,49 ) were prepared from two previously described glycan libraries 20,48,49 ). Amine containing glycans with spacer’s sp2, sp3 or sp4 were synthesised as previously described 20 and glycans without spacers were amine functionalised as previously published 50 .…”

Section: Methodsmentioning

confidence: 99%

YesU from Bacillus subtilis preferentially binds fucosylated glycans

Tiralongo

Cooper

Litfin

et al. 2018

Sci Rep

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The interaction of carbohydrate-binding proteins (CBPs) with their corresponding glycan ligands is challenging to study both experimentally and computationally. This is in part due to their low binding affinity, high flexibility, and the lack of a linear sequence in carbohydrates, as exists in nucleic acids and proteins. We recently described a function-prediction technique called SPOT-Struc that identifies CBPs by global structural alignment and binding-affinity prediction. Here we experimentally determined the carbohydrate specificity and binding affinity of YesU (RCSB PDB ID: 1oq1), an uncharacterized protein from Bacillus subtilis that SPOT-Struc predicted would bind high mannose-type glycans. Glycan array analyses however revealed glycan binding patterns similar to those exhibited by fucose (Fuc)-binding lectins, with SPR analysis revealing high affinity binding to Lewisx and lacto-N-fucopentaose III. Structure based alignment of YesU revealed high similarity to the legume lectins UEA-I and GS-IV, and docking of Lewisx into YesU revealed a complex structure model with predicted binding affinity of −4.3 kcal/mol. Moreover the adherence of B. subtilis to intestinal cells was significantly inhibited by Lex and Ley but by not non-fucosylated glycans, suggesting the interaction of YesU to fucosylated glycans may be involved in the adhesion of B. subtilis to the gastrointestinal tract of mammals.

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Differential carbohydrate binding and cell surface glycosylation of human cancer cell lines

Cited by 53 publications

References 27 publications

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Identification of NuoX and NuoY Ligand Binding Specificity in the Campylobacter Jejuni Complex I

YesU from Bacillus subtilis preferentially binds fucosylated glycans

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