Tryptophan fluorescence properties of cholera toxin upon interacting with ganglioside GD<sub>1b</sub>

Mestrallet, Marta Graciela; Bennun, Fernando Ruben; Maggio, Bruno; Cumar, Federico A.

doi:10.1002/jnr.490120220

Cited by 7 publications

(2 citation statements)

References 25 publications

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“…The appearance of signal corresponding to GD1a/b is consistent with the results of previously reported binding experiments that identified GD1b as a low affinity ligand for CTB 5 . 34,40,41 Direct ESI-MS binding measurements performed on solutions of CTB 5 with GD1a or GD1b revealed that GD1b binds with a measurable affinity, while GD1a does not bind (data not shown).…”

Section: ■ Results and Discussionmentioning

confidence: 98%

Nanodiscs and Electrospray Ionization Mass Spectrometry: A Tool for Screening Glycolipids Against Proteins

et al. 2014

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Electrospray ionization-mass spectrometry (ESI-MS) is extensively employed to detect and quantify protein-carbohydrate interactions in vitro and is increasingly used to screen carbohydrate libraries against target proteins. However, current ESI-MS methods are limited to carbohydrate ligands that are relatively soluble in water and are, therefore, not generally suitable for studying protein interactions with glycolipids, an important class of cellular receptors. Here, we describe a catch-and-release (CaR)-ESI-MS assay, which exploits nanodiscs (NDs) to solubilize glycolipids and mimic their natural cellular environment, suitable for screening libraries of glycosphingolipids (GSL) against proteins to identify specific interactions and to rank their relative affinities. Using the B subunit homopentamers of cholera toxin and heat labile toxin as model GSL-binding proteins, the CaR-ESI-MS was applied to NDs containing mixtures of gangliosides. The results demonstrate that the CaR-ESI-MS assay can simultaneously detect both high and low affinity GSL ligands using either a library of NDs that each contains one GSL or incorporating a mixture of GSLs into a single ND. Moreover, the relative abundances of the released ligands appear to reflect their relative affinities in solution. Application of the CaR-ESI-MS assay using NDs containing gangliosides extracted from porcine brain led to the discovery of a neolacto GSL as a cholera toxin ligand, highlighting the power of the assay for identifying specific protein-glycolipid interactions from biologically relevant mixtures.

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Section: ■ Results and Discussionmentioning

confidence: 98%

Nanodiscs and Electrospray Ionization Mass Spectrometry: A Tool for Screening Glycolipids Against Proteins

et al. 2014

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“…It is generally accepted that the monosialoganglioside GM1 is the toxin receptor in the intestine of several animal species including humans (6,16,19). Recently, it has been shown that other gangliosides such as GDlb (8,15,23), fucose-GM1 (18), and galactose-N-acetylgalactosamine-GM1l (26) show considerable capacity to bind CT when assayed in vitro. Even if some of these latter gangliosides are also present in intestinal epithelial cells, a receptor capability leading to an increase in cyclic AMP concentration has not yet been demonstrated for any of them.…”

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Binding of cholera toxin to pig intestinal mucosa glycosphingolipids: relationship with the ABO blood group system

et al. 1989

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A search for compounds from intestinal mucosa of pigs carrying and not carrying blood group A-active substances (A+ and A-pigs, respectively) capable of binding cholera toxin (CT) was performed. Glycolipid extracts from a pool of pig intestinal mucosa resolved in thin-layer chromatography (TLC) revealed the presence of six to eight compounds capable of binding 12'I-CT, two of them running as the ganglioside standards GM, and GDlb. When intestinal mucosa glycolipids from single pigs were assayed by TLC for CTbinding capacity, two different patterns of labeling were observed. The main difference was at the level of compounds running below GDlb. The A' pigs but not the A-pigs showed CT binding at this level. The major CT-binding compound detected only in A+ pigs was purified and some properties were determined. After TLC developed with different solvent systems, the purified compound bound CT and also immunoreacted with anti-A and anti-AB antisera but not with anti-B antiserum. The compound was also able to inhibit the hemagglutination of human A erythrocytes caused by anti-A antiserum, but inhibition was not observed with the B-anti-B or 0 (H)-Ulex europaeus lectin systems. A partial chemical characterization indicated that the active compound is a neutral glycosphingolipid containing glucose, fucose, galactose, and hexosamine. The existence of a blood group-active substance(s) able to interact with CT may help to explain the relationship between ABO blood groups and the diarrheal disease caused by infection with Vibrio cholerae.

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Escherichia coli heat-labile enterotoxin preferentially interacts with blood group A-active glycolipids from pig intestinal mucosa and A- and B-active glycolipids from human red cells compared to H-active glycolipids

et al. 1992

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The capacity of cholera toxin (CT) and of the heat-labile enterotoxin produced by Escherichia coli isolated from humans (LTh) to interact with glycolipids bearing ABO(H) blood group determinants isolated from different sources and separated by thin layer chromatography was studied. Toxin binding to the ABO(H)-related glycolipids depends on the glycolipid source, the type of the blood group activity, and the toxin. LTh and CT were capable of interacting with several blood group-active glycolipids from pig intestinal mucosa and both toxins preferentially recognize glycolipids isolated from animals carrying A-blood group antigenic determinants compared to those isolated from animals lacking these antigens. In contrast, LTh but not CT was able to interact with ABO(H)-active glycolipids from human erythrocytes. LTh preferentially binds to glycolipids isolated from A, B, and AB compared to O red cells. Results from competition experiments between CT and LTh for binding to the blood group-active glycolipids suggest that the carbohydrate structure requirements for the interaction of each toxin are different. The present findings may help to understand the results of clinical studies indicating an association between ABO(H) blood groups and the severity of diarrheal diseases produced by some toxigenic enterobacteria.

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Tryptophan fluorescence properties of cholera toxin upon interacting with ganglioside GD_1b

Cited by 7 publications

References 25 publications

Nanodiscs and Electrospray Ionization Mass Spectrometry: A Tool for Screening Glycolipids Against Proteins

Nanodiscs and Electrospray Ionization Mass Spectrometry: A Tool for Screening Glycolipids Against Proteins

Binding of cholera toxin to pig intestinal mucosa glycosphingolipids: relationship with the ABO blood group system

Escherichia coli heat-labile enterotoxin preferentially interacts with blood group A-active glycolipids from pig intestinal mucosa and A- and B-active glycolipids from human red cells compared to H-active glycolipids

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Tryptophan fluorescence properties of cholera toxin upon interacting with ganglioside GD1b

Cited by 7 publications

References 25 publications

Nanodiscs and Electrospray Ionization Mass Spectrometry: A Tool for Screening Glycolipids Against Proteins

Nanodiscs and Electrospray Ionization Mass Spectrometry: A Tool for Screening Glycolipids Against Proteins

Binding of cholera toxin to pig intestinal mucosa glycosphingolipids: relationship with the ABO blood group system

Escherichia coli heat-labile enterotoxin preferentially interacts with blood group A-active glycolipids from pig intestinal mucosa and A- and B-active glycolipids from human red cells compared to H-active glycolipids

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Tryptophan fluorescence properties of cholera toxin upon interacting with ganglioside GD_1b