Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1.

L. O. Sillerud, J. H. Prestegard, Robert K Yu, W. H. Konigsberg, D. E. Schafer

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The initial event in the action of cholera toxin on intact cells is its recognition of cell-surface receptors, molecules of ganglioside GM1. We have studied details of this interaction by 13C NMR, which enables us to examine simultaneously both the protein and the ganglioside or, as in the present instance, its oligosaccharide portion. 13C NMR spectra of the toxin are consistent with the long correlation times expected for this 84,000-dalton protein. They show, however, some resolved resonances (including one tentatively assigned to the epsilon 2 carbon of tryptophan, 138.3 ppm downfield from tetramethylsilane). When oligosaccharide is added to the toxin this resonance broadens or moves further upfield to reside under phenylalanine resonances at 136.7 ppm. Of the seven tryptophan residues in cholera toxin, five are in the B subunits which bind GM1, so that the data are consistent with a resonance shift for the epsilon 2 carbons of these residues on binding the oligosaccharide. Resonances arising from the anomeric and methylene carbons of the sialic acid moiety of the oligosaccharide are also shifted. Comparison with corresponding chemical shifts in a series of model compounds suggest that the latter effects may originate in a toxin-induced conformational change in the oligosaccharide. At high resolution, anomeric carbon resonances of terminal galactose residues in free and bound oligosaccharide are also resolved.

Original languageEnglish (US)
Pages (from-to)1094-1097
Number of pages4
JournalJournal of Biological Chemistry
Volume256
Issue number3
StatePublished - Feb 10 1981
Externally publishedYes

Fingerprint

G(M1) Ganglioside
Cholera Toxin
Oligosaccharides
Nuclear magnetic resonance
Observation
Carbon
Tryptophan
Gangliosides
Chemical shift
Cell Surface Receptors
N-Acetylneuraminic Acid
Phenylalanine
Galactose
Carbon-13 Magnetic Resonance Spectroscopy
Proteins
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sillerud, L. O., Prestegard, J. H., Yu, R. K., Konigsberg, W. H., & Schafer, D. E. (1981). Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1. Journal of Biological Chemistry, 256(3), 1094-1097.

Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1. / Sillerud, L. O.; Prestegard, J. H.; Yu, Robert K; Konigsberg, W. H.; Schafer, D. E.

In: Journal of Biological Chemistry, Vol. 256, No. 3, 10.02.1981, p. 1094-1097.

Research output: Contribution to journalArticle

Sillerud, LO, Prestegard, JH, Yu, RK, Konigsberg, WH & Schafer, DE 1981, 'Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1.', Journal of Biological Chemistry, vol. 256, no. 3, pp. 1094-1097.
Sillerud, L. O. ; Prestegard, J. H. ; Yu, Robert K ; Konigsberg, W. H. ; Schafer, D. E. / Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1. In: Journal of Biological Chemistry. 1981 ; Vol. 256, No. 3. pp. 1094-1097.
@article{13800eff395446efb3db8ab8f63a38a1,
title = "Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1.",
abstract = "The initial event in the action of cholera toxin on intact cells is its recognition of cell-surface receptors, molecules of ganglioside GM1. We have studied details of this interaction by 13C NMR, which enables us to examine simultaneously both the protein and the ganglioside or, as in the present instance, its oligosaccharide portion. 13C NMR spectra of the toxin are consistent with the long correlation times expected for this 84,000-dalton protein. They show, however, some resolved resonances (including one tentatively assigned to the epsilon 2 carbon of tryptophan, 138.3 ppm downfield from tetramethylsilane). When oligosaccharide is added to the toxin this resonance broadens or moves further upfield to reside under phenylalanine resonances at 136.7 ppm. Of the seven tryptophan residues in cholera toxin, five are in the B subunits which bind GM1, so that the data are consistent with a resonance shift for the epsilon 2 carbons of these residues on binding the oligosaccharide. Resonances arising from the anomeric and methylene carbons of the sialic acid moiety of the oligosaccharide are also shifted. Comparison with corresponding chemical shifts in a series of model compounds suggest that the latter effects may originate in a toxin-induced conformational change in the oligosaccharide. At high resolution, anomeric carbon resonances of terminal galactose residues in free and bound oligosaccharide are also resolved.",
author = "Sillerud, {L. O.} and Prestegard, {J. H.} and Yu, {Robert K} and Konigsberg, {W. H.} and Schafer, {D. E.}",
year = "1981",
month = "2",
day = "10",
language = "English (US)",
volume = "256",
pages = "1094--1097",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Observation by 13C NMR of interactions between cholera toxin and the oligosaccharide of ganglioside GM1.

AU - Sillerud, L. O.

AU - Prestegard, J. H.

AU - Yu, Robert K

AU - Konigsberg, W. H.

AU - Schafer, D. E.

PY - 1981/2/10

Y1 - 1981/2/10

N2 - The initial event in the action of cholera toxin on intact cells is its recognition of cell-surface receptors, molecules of ganglioside GM1. We have studied details of this interaction by 13C NMR, which enables us to examine simultaneously both the protein and the ganglioside or, as in the present instance, its oligosaccharide portion. 13C NMR spectra of the toxin are consistent with the long correlation times expected for this 84,000-dalton protein. They show, however, some resolved resonances (including one tentatively assigned to the epsilon 2 carbon of tryptophan, 138.3 ppm downfield from tetramethylsilane). When oligosaccharide is added to the toxin this resonance broadens or moves further upfield to reside under phenylalanine resonances at 136.7 ppm. Of the seven tryptophan residues in cholera toxin, five are in the B subunits which bind GM1, so that the data are consistent with a resonance shift for the epsilon 2 carbons of these residues on binding the oligosaccharide. Resonances arising from the anomeric and methylene carbons of the sialic acid moiety of the oligosaccharide are also shifted. Comparison with corresponding chemical shifts in a series of model compounds suggest that the latter effects may originate in a toxin-induced conformational change in the oligosaccharide. At high resolution, anomeric carbon resonances of terminal galactose residues in free and bound oligosaccharide are also resolved.

AB - The initial event in the action of cholera toxin on intact cells is its recognition of cell-surface receptors, molecules of ganglioside GM1. We have studied details of this interaction by 13C NMR, which enables us to examine simultaneously both the protein and the ganglioside or, as in the present instance, its oligosaccharide portion. 13C NMR spectra of the toxin are consistent with the long correlation times expected for this 84,000-dalton protein. They show, however, some resolved resonances (including one tentatively assigned to the epsilon 2 carbon of tryptophan, 138.3 ppm downfield from tetramethylsilane). When oligosaccharide is added to the toxin this resonance broadens or moves further upfield to reside under phenylalanine resonances at 136.7 ppm. Of the seven tryptophan residues in cholera toxin, five are in the B subunits which bind GM1, so that the data are consistent with a resonance shift for the epsilon 2 carbons of these residues on binding the oligosaccharide. Resonances arising from the anomeric and methylene carbons of the sialic acid moiety of the oligosaccharide are also shifted. Comparison with corresponding chemical shifts in a series of model compounds suggest that the latter effects may originate in a toxin-induced conformational change in the oligosaccharide. At high resolution, anomeric carbon resonances of terminal galactose residues in free and bound oligosaccharide are also resolved.

UR - http://www.scopus.com/inward/record.url?scp=0019876629&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0019876629&partnerID=8YFLogxK

M3 - Article

VL - 256

SP - 1094

EP - 1097

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -