WaterLOGSY as a method for primary NMR screening: practical aspects and range of applicability

• Published in: Journal of biomolecular NMR Vol. 21; no. 4; pp. 349 - 359
• Main Authors: Dalvit, C, Fogliatto, G, Stewart, A, Veronesi, M, Stockman, B
• Format: Journal Article
• Language: English
• Published: Netherlands Springer Nature B.V 01-12-2001
• Subjects: Binding, Competitive; CDC2-CDC28 Kinases; Chemistry, Pharmaceutical - methods; Combinatorial Chemistry Techniques; Cyclin A - chemistry; Cyclin A - metabolism; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinases - chemistry; Cyclin-Dependent Kinases - metabolism; Drug Evaluation, Preclinical - methods; Glutathione Transferase - metabolism; Humans; Ligands; Nuclear Magnetic Resonance, Biomolecular - methods; Protein Binding; Protein-Serine-Threonine Kinases - chemistry; Protein-Serine-Threonine Kinases - metabolism; Sensitivity and Specificity; Structure-Activity Relationship; Time Factors; Water - chemistry
• ISSN: 0925-2738; 1573-5001
• DOI: 10.1023/a:1013302231549

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.