TINS, target immobilized NMR screening: an efficient and sensitive method for ligand discovery.
We propose a ligand screening method, called TINS (target immobilized NMR screening), which reduces the amount of target required for the fragment-based approach to drug discovery. Binding is detected by comparing 1D NMR spectra of compound mixtures in the presence of a target immobilized on a solid support to a control sample. The method has been validated by the detection of a variety of ligands for protein and nucleic acid targets (K(D) from 60 to 5000 muM). The ligand binding capacity of a protein was undiminished after 2000 different compounds had been applied, indicating the potential to apply the assay for screening typical fragment libraries. TINS can be used in competition mode, allowing rapid characterization of the ligand binding site. TINS may allow screening of targets that are difficult to produce or that are insoluble, such as membrane proteins.
Vanwetswinkel
Sophie
S
Leiden Institute of Chemistry, Leiden University, Postbus 9502, 2300-RA Leiden, The Netherlands.
Heetebrij
Robert J
RJ
van Duynhoven
John
J
Hollander
Johan G
JG
Filippov
Dmitri V
DV
Hajduk
Philip J
PJ
Siegal
Gregg
G
D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000737:chemistry*; D007202:Indicators and Reagents; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D012680:Sensitivity and Specificity; D022021:Tacrolimus Binding Proteins / Q000737:chemistry
D007202:Indicators and Reagents; D008024:Ligands; D009841:Oligonucleotides; D011506:Proteins; D012313:RNA; D004247:DNA; D022021:Tacrolimus Binding Proteins