Transferred nuclear Overhauser enhancement (NOE) and rotating-frame NOE experiments reflect the size of the bound segment of the Forssman pentasaccharide in the binding site of Dolichos biflorus lectin

Florence Casset, Anne Imberty, Serge Pérez, Marilynn E. Etzler, Hans Paulsen, Thomas Peters*

*Korrespondierende/r Autor/-in für diese Arbeit
13 Zitate (Scopus)

Abstract

A complex between the Forssman pentasaccharide α-D-GalNAc-(1→3)-β-D-GalNAc-(1→3)-α-D-Gal-(1→4)-β-D-Gal-(1→4)-D-Glc and the seed lectin from Dolichos biflorus was studied using transfer-NOESY and transfer rotating frame NOE spectroscopy (ROESY) experiments. The evolution of transferred NOEs and ROEs as a function of the pentasaccharide/lectin ratio was different for the non-reducing disaccharide moiety α-D-GalNAc-(1→3)-β-D-GalNAc compared to the rest of the molecule, which reflects distinct relaxation properties and effects of exchange broadening of the corresponding ligand resonances. Significantly, several intermolecular transferred NOEs were observed between protons of the non-reducing disaccharide moiety α-D-GalNAc-(1→3)-β-D-GalNAc and aliphatic as well as aromatic amino acid side chain protons in the binding pocket of the lectin. It is concluded that the non-reducing disaccharide fragment is buried in the lectin-binding pocket, whereas the reducing trisaccharide portion α-D-Gal-(1→4)-β-D-Gal-(1→4)-D-Glc has no immediate contacts with the protein. The experimental transfer NOE data were qualitatively compared to theoretical proton-proton distances from a model that was based on a previous homology modeling study of a complex between the disaccharide fragment α-D-GalNAc-(1→3)-β-D-GalNAc and D. biflorus lectin. It was found that all intermolecular transferred NOEs matched short interatomic distances between ligand protons and aliphatic or aromatic amino acid side chain protons predicted by the theoretical model.

OriginalspracheEnglisch
ZeitschriftEuropean Journal of Biochemistry
Jahrgang244
Ausgabenummer1
Seiten (von - bis)242-250
Seitenumfang9
ISSN0014-2956
DOIs
PublikationsstatusVeröffentlicht - 01.01.1997

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