title: Intermolecular interaction studies using small volumes.
abstract: 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.
authors.affiliation: Department of Chemistry, Ghent University, Ghent, Belgium.
authors.forename: David
authors.initials: D
authors.lastname: Bourry
authors.forename: Davy
authors.initials: D
authors.lastname: Sinnaeve
authors.forename: Katelijne
authors.initials: K
authors.lastname: Gheysen
authors.forename: Bernd
authors.initials: B
authors.lastname: Fritzinger
authors.forename: Gianni
authors.initials: G
authors.lastname: Vandenborre
authors.forename: Els J M
authors.initials: EJ
authors.lastname: Van Damme
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Wieruszeski
authors.forename: Guy
authors.initials: G
authors.lastname: Lippens
authors.forename: Christophe
authors.initials: C
authors.lastname: Ampe
authors.forename: José C
authors.initials: JC
authors.lastname: Martins
mesh_terms: D006801:Humans; D037102:Lectins / Q000737:chemistry*; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000592:standards; D015394:Molecular Structure; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D051304:Profilins / Q000737:chemistry*; D012015:Reference Standards; D013696:Temperature
chemical_list: D037102:Lectins; D008024:Ligands; D019269:Phosphatidylinositol 4,5-Diphosphate; D011134:Polysaccharides; D051304:Profilins