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Intermolecular interaction studies using small volumes
Author(s) -
Bourry David,
Sinnaeve Davy,
Gheysen Katelijne,
Fritzinger Bernd,
Vandenborre Gianni,
Van Damme Els J. M.,
Wieruszeski JeanMichel,
Lippens Guy,
Ampe Christophe,
Martins José C.
Publication year - 2011
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.2699
Subject(s) - chemistry , titration , intermolecular force , ligand (biochemistry) , analytical chemistry (journal) , volume (thermodynamics) , chemical physics , molecule , thermodynamics , chromatography , organic chemistry , biochemistry , physics , receptor
We present the use of 1‐mm room‐temperature probe technology to perform intermolecular interaction studies using chemical shift perturbation methods and saturation transfer difference (STD) spectroscopy using small sample volumes. The use of a small sample volume (5–10 µl) allows for an alternative titration protocol where individual samples are prepared for each titration point, rather than the usual protocol used for a 5‐mm probe setup where the ligand is added consecutively to the solution containing the protein or host of interest. This allows for considerable economy in the consumption and cost of the protein and ligand amounts required for interaction studies. For titration experiments, the use of the 1‐mm setup consumes less than 10% of the ligand amount required using a 5‐mm setup. This is especially significant when complex ligands that are only available in limited quantities, typically because they are obtained from natural sources or through elaborate synthesis efforts, need to be investigated. While the use of smaller volumes does increase the measuring time, we demonstrate that the use of commercial small volume probes allows the study of interactions that would otherwise be impossible to address by NMR. Copyright © 2010 John Wiley & Sons, Ltd.