Application of homonuclear 3D NMR experiments and 1D analogs to study the conformation of sialyl Lewisx bound to E-selectin

Karoline Scheffler; Jean Robert Brisson; Rüdiger Weisemann; John L. Magnani; Wei Tong Wong; Beat Ernst; Thomas Peters

doi:10.1023/A:1018358929268

Application of homonuclear 3D NMR experiments and 1D analogs to study the conformation of sialyl Lewis^x bound to E-selectin

Karoline Scheffler, Jean Robert Brisson, Rüdiger Weisemann, John L. Magnani, Wei Tong Wong, Beat Ernst, Thomas Peters^*

^*Corresponding author for this work

Institute of Chemistry and Metabolomics

45 Citations (Scopus)

Abstract

The conformation of the sialyl Lewis^x tetrasaccharide bound to E-selectin was previously determined from transfer NOE (trNOE) experiments in conjunction with a distance-geometry analysis. However, the orientation of the tetrasaccharide ligand in the binding site of E-selectin is still unknown. It can be predicted that the accurate quantitative analysis of all trNOEs, including those originating from spin diffusion, is one key to analyze the orientation of sialyl Lewis^x in the binding pocket of E-selectin. Therefore, we applied homonuclear 3D NMR experiments and 1D analogs to obtain trNOEs that could not unambiguously be assigned from previous 2D trNOESY spectra, due to severe resonance-signal overlap. A 3D TOCSY-trNOESY experiment, a 1D TOCSY-trNOESY experiment, and a 1D trNOESY-TOCSY experiment of the sialyl Lewis^x/E-selectin complex furnished new interglycosidic trNOEs and provided additional information for the interpretation of trNOEs that have been described before. A 2D trROESY spectrum of the sialyl Lewis^x/E-selectin complex allowed one to identify the amount of spin-diffusion contributions to trNOEs. Finally, an unambiguous assignment of all trNOEs, and an analysis of spin-diffusion pathways, was obtained, creating a basis for a quantitative analysis of trNOEs in the sialyl Lewis^x/E-selectin complex.

Original language	English
Journal	Journal of Biomolecular NMR
Volume	9
Issue number	4
Pages (from-to)	423-436
Number of pages	14
ISSN	0925-2738
DOIs	https://doi.org/10.1023/A:1018358929268
Publication status	Published - 1997

Funding

This work was supported by a grant of the Deutsche Forschungsgemeinschaft and by the Fonds der Chemi-schen Industrie. T.P. and J.-R.B. wish to thank NATO for a travel grant (CRG 890356). We are indebted to Prof. Dr. H. Rüterjans (Institute for Biophysical Chemistry, University of Frankfurt, Frankfurt, Germany) for giving us access to the 600 MHz spectrometer in his laboratory.

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1023/A:1018358929268

Cite this

@article{9e5b96d5bd1a431b8b9c752246fe9e1a,

title = "Application of homonuclear 3D NMR experiments and 1D analogs to study the conformation of sialyl Lewisx bound to E-selectin",

abstract = "The conformation of the sialyl Lewisx tetrasaccharide bound to E-selectin was previously determined from transfer NOE (trNOE) experiments in conjunction with a distance-geometry analysis. However, the orientation of the tetrasaccharide ligand in the binding site of E-selectin is still unknown. It can be predicted that the accurate quantitative analysis of all trNOEs, including those originating from spin diffusion, is one key to analyze the orientation of sialyl Lewisx in the binding pocket of E-selectin. Therefore, we applied homonuclear 3D NMR experiments and 1D analogs to obtain trNOEs that could not unambiguously be assigned from previous 2D trNOESY spectra, due to severe resonance-signal overlap. A 3D TOCSY-trNOESY experiment, a 1D TOCSY-trNOESY experiment, and a 1D trNOESY-TOCSY experiment of the sialyl Lewisx/E-selectin complex furnished new interglycosidic trNOEs and provided additional information for the interpretation of trNOEs that have been described before. A 2D trROESY spectrum of the sialyl Lewisx/E-selectin complex allowed one to identify the amount of spin-diffusion contributions to trNOEs. Finally, an unambiguous assignment of all trNOEs, and an analysis of spin-diffusion pathways, was obtained, creating a basis for a quantitative analysis of trNOEs in the sialyl Lewisx/E-selectin complex.",

author = "Karoline Scheffler and Brisson, \{Jean Robert\} and R{\"u}diger Weisemann and Magnani, \{John L.\} and Wong, \{Wei Tong\} and Beat Ernst and Thomas Peters",

note = "Funding Information: This work was supported by a grant of the Deutsche Forschungsgemeinschaft and by the Fonds der Chemi-schen Industrie. T.P. and J.-R.B. wish to thank NATO for a travel grant (CRG 890356). We are indebted to Prof. Dr. H. R{\"u}terjans (Institute for Biophysical Chemistry, University of Frankfurt, Frankfurt, Germany) for giving us access to the 600 MHz spectrometer in his laboratory. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.",

year = "1997",

doi = "10.1023/A:1018358929268",

language = "English",

volume = "9",

pages = "423--436",

journal = "Journal of Biomolecular NMR",

issn = "0925-2738",

publisher = "Springer Science and Business Media B.V.",

number = "4",

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TY - JOUR

T1 - Application of homonuclear 3D NMR experiments and 1D analogs to study the conformation of sialyl Lewisx bound to E-selectin

AU - Scheffler, Karoline

AU - Brisson, Jean Robert

AU - Weisemann, Rüdiger

AU - Magnani, John L.

AU - Wong, Wei Tong

AU - Ernst, Beat

AU - Peters, Thomas

N1 - Funding Information: This work was supported by a grant of the Deutsche Forschungsgemeinschaft and by the Fonds der Chemi-schen Industrie. T.P. and J.-R.B. wish to thank NATO for a travel grant (CRG 890356). We are indebted to Prof. Dr. H. Rüterjans (Institute for Biophysical Chemistry, University of Frankfurt, Frankfurt, Germany) for giving us access to the 600 MHz spectrometer in his laboratory. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.

PY - 1997

Y1 - 1997

N2 - The conformation of the sialyl Lewisx tetrasaccharide bound to E-selectin was previously determined from transfer NOE (trNOE) experiments in conjunction with a distance-geometry analysis. However, the orientation of the tetrasaccharide ligand in the binding site of E-selectin is still unknown. It can be predicted that the accurate quantitative analysis of all trNOEs, including those originating from spin diffusion, is one key to analyze the orientation of sialyl Lewisx in the binding pocket of E-selectin. Therefore, we applied homonuclear 3D NMR experiments and 1D analogs to obtain trNOEs that could not unambiguously be assigned from previous 2D trNOESY spectra, due to severe resonance-signal overlap. A 3D TOCSY-trNOESY experiment, a 1D TOCSY-trNOESY experiment, and a 1D trNOESY-TOCSY experiment of the sialyl Lewisx/E-selectin complex furnished new interglycosidic trNOEs and provided additional information for the interpretation of trNOEs that have been described before. A 2D trROESY spectrum of the sialyl Lewisx/E-selectin complex allowed one to identify the amount of spin-diffusion contributions to trNOEs. Finally, an unambiguous assignment of all trNOEs, and an analysis of spin-diffusion pathways, was obtained, creating a basis for a quantitative analysis of trNOEs in the sialyl Lewisx/E-selectin complex.

AB - The conformation of the sialyl Lewisx tetrasaccharide bound to E-selectin was previously determined from transfer NOE (trNOE) experiments in conjunction with a distance-geometry analysis. However, the orientation of the tetrasaccharide ligand in the binding site of E-selectin is still unknown. It can be predicted that the accurate quantitative analysis of all trNOEs, including those originating from spin diffusion, is one key to analyze the orientation of sialyl Lewisx in the binding pocket of E-selectin. Therefore, we applied homonuclear 3D NMR experiments and 1D analogs to obtain trNOEs that could not unambiguously be assigned from previous 2D trNOESY spectra, due to severe resonance-signal overlap. A 3D TOCSY-trNOESY experiment, a 1D TOCSY-trNOESY experiment, and a 1D trNOESY-TOCSY experiment of the sialyl Lewisx/E-selectin complex furnished new interglycosidic trNOEs and provided additional information for the interpretation of trNOEs that have been described before. A 2D trROESY spectrum of the sialyl Lewisx/E-selectin complex allowed one to identify the amount of spin-diffusion contributions to trNOEs. Finally, an unambiguous assignment of all trNOEs, and an analysis of spin-diffusion pathways, was obtained, creating a basis for a quantitative analysis of trNOEs in the sialyl Lewisx/E-selectin complex.

UR - https://www.scopus.com/pages/publications/0031157117

U2 - 10.1023/A:1018358929268

DO - 10.1023/A:1018358929268

M3 - Journal articles

C2 - 9255946

AN - SCOPUS:0031157117

SN - 0925-2738

VL - 9

SP - 423

EP - 436

JO - Journal of Biomolecular NMR

JF - Journal of Biomolecular NMR

IS - 4

ER -