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Saturation transfer double‐difference NMR spectroscopy using a dual solenoid microcoil difference probe
Author(s) -
Bergeron Scott J.,
Henry Ian D.,
Santini Robert E.,
Aghdasi Abdollah,
Raftery Daniel
Publication year - 2008
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/mrc.2275
Subject(s) - chemistry , microcoil , pulse sequence , analytical chemistry (journal) , proton nmr , nuclear magnetic resonance spectroscopy , nuclear magnetic resonance , ligand (biochemistry) , solenoid , electromagnetic coil , stereochemistry , chromatography , receptor , physics , electrical engineering , engineering , mechanical engineering , biochemistry
An experiment designed to collect a saturation transfer double difference (STDD) NMR spectrum using a solenoid microcoil NMR difference probe is reported. STDD‐NMR allows the investigation of ligand‐biomolecule binding, with moderate concentration requirements for unlabeled molecular targets and the ability to discern binding events in the presence of non‐binding ligands. The NMR difference probe acquires the signals from two different samples at once, and cancels common signals automatically through a mechanism of switching between parallel excitation and serial acquisition of the sample signals. STDD spectra were acquired on a system consisting of human serum albumin and two ligands, octanoic acid and glucose. The non‐binding ligand, glucose, was cancelled internally through phase cycling, while the protein signal was subtracted automatically by the difference probe. The proton NMR resonance signal from octanoic acid remained in the double difference spectrum. This work demonstrates that the double difference can be performed both internally and automatically through the utilization of the solenoid microcoil NMR difference probe and STDD‐NMR pulse sequence, resulting in a clean signal from the binding ligand with good protein background subtraction and an overall favorable result when compared to the conventional approach. Copyright © 2008 John Wiley & Sons, Ltd.