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Extra resources for Biophysical approaches determining ligand binding to biomolecular targets : detection, measurement and modelling
Due to the transient nature of the NOE transfer in the mixing time of a NOESY pulse sequence, this method is less sensitive than STD or WaterLOGSY methods. However, tr-NOE remains an appealing method due to its simplicity. Moreover, since positive 35 Nuclear Magnetic Resonance of Ligand Binding to Proteins NOE cross-peaks that are observed for a binding ligand are speciﬁc to its bound conformation, their analysis in terms of inter-proton distances may be used to get structural information about the bound conformation of the ligand.
The large molecular receptor is polarised using a selective pulse on one of the receptor resonances (often a methyl group) at a frequency where no ligand resonance is found. The saturation (shown in blue) is transferred on the binding ligand altering the intensity of its resonance lines, which allows its identiﬁcation by subtracting spectra recorded with and without selective irradiation of the receptor. Due to its large size (that translates in broad resonance peaks in the spectrum) and small quantity (ligands are in large excess), the receptor is not visible in the spectrum.
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