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Fluorinated Carbohydrates as Lectin Ligands: Dissecting Glycan–Cyanovirin Interactions by Using 19 F NMR Spectroscopy
Author(s) -
Matei Elena,
André Sabine,
Glinschert Anja,
Infantino Angela Simona,
Oscarson Stefan,
Gabius HansJoachim,
Gronenborn Angela M.
Publication year - 2013
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201204070
Subject(s) - isothermal titration calorimetry , chemistry , nuclear magnetic resonance spectroscopy , heteronuclear single quantum coherence spectroscopy , glycan , lectin , fluorine 19 nmr , ligand (biochemistry) , spectroscopy , titration , two dimensional nuclear magnetic resonance spectroscopy , stereochemistry , chemical shift , biochemistry , organic chemistry , glycoprotein , physics , receptor , quantum mechanics
Abstract NMR spectroscopy and isothermal titration calorimetry (ITC) are powerful methods to investigate ligand–protein interactions. Here, we present a versatile and sensitive fluorine NMR spectroscopic approach that exploits the 19 F nucleus of 19 F‐labeled carbohydrates as a sensor to study glycan binding to lectins. Our approach is illustrated with the 11 kDa Cyanovirin‐N, a mannose binding anti‐HIV lectin. Two fluoro‐deoxy sugar derivatives, methyl 2‐deoxy‐2‐fluoro‐α‐ D ‐mannopyranosyl‐(1→2)‐α‐ D ‐mannopyranoside and methyl 2‐deoxy‐2‐fluoro‐α‐ D ‐mannopyranosyl‐(1→2)‐α‐ D ‐mannopyranosyl‐(1→2)‐α‐ D ‐mannopyranoside were utilized. Binding was studied by 19 F NMR spectroscopy of the ligand and 1 H– 15 N HSQC NMR spectroscopy of the protein. The NMR data agree well with those obtained from the equivalent reciprocal and direct ITC titrations. Our study shows that the strategic design of fluorinated ligands and fluorine NMR spectroscopy for ligand screening holds great promise for easy and fast identification of glycan binding, as well as for their use in reporting structural and/or electronic perturbations that ensue upon interaction with a cognate lectin.