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 - http://www.scopus.com/inward/record.url?scp=0031157117&partnerID=8YFLogxK
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 -
