title: WaterLOGSY as a method for primary NMR screening: practical aspects and range of applicability.
abstract: WaterLOGSY represents a powerful method for primary NMR screening in the identification of compounds interacting with macromolecules, including proteins and DNA or RNA fragments. Several relay pathways are used constructively in the experiment for transferring bulk water magnetization to the ligand. The method is particularly useful for the identification of novel scaffolds of micromolar affinity that can be then optimized using directed screening, combinatorial chemistry, medicinal chemistry and structure-based drug design. The practical aspects and range of applicability of the WaterLOGSY experiment are analyzed in detail here. Competition binding and titration WaterLOGSY permit, after proper correction, the evaluation of the dissociation binding constant. The high sensitivity of the technique in combination with the easy deconvolution of the mixtures for the identification of the active components, significantly reduces the amount of material and time needed for the NMR screening process.
authors.affiliation: Chemistry Department, Pharmacia, Nerviano, Italy. claudio.dalvit@eu.pnu.com
authors.forename: C
authors.initials: C
authors.lastname: Dalvit
authors.forename: G
authors.initials: G
authors.lastname: Fogliatto
authors.forename: A
authors.initials: A
authors.lastname: Stewart
authors.forename: M
authors.initials: M
authors.lastname: Veronesi
authors.forename: B
authors.initials: B
authors.lastname: Stockman
mesh_terms: D001667:Binding, Competitive; D042846:CDC2-CDC28 Kinases; D002626:Chemistry, Pharmaceutical / Q000379:methods; D020650:Combinatorial Chemistry Techniques; D019925:Cyclin A / Q000737:chemistry / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000737:chemistry / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008024:Ligands; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship; D013997:Time Factors; D014867:Water / Q000737:chemistry*
chemical_list: D019925:Cyclin A; D008024:Ligands; D014867:Water; D005982:Glutathione Transferase; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases