Labeled Ligand Displacement: Extending NMR-Based Screening of Protein Targets.

NMR spectroscopy has enjoyed widespread success as a method for screening protein targets, especially in the area of fragment-based drug discovery. However, current methods for NMR-based screening all suffer certain limitations. Two-dimensional methods like "SAR by NMR" require isotopically labeled protein and are limited to proteins less than about 50 kDa. For one-dimensional, ligand-based methods, results can be confounded by nonspecific compound binding, resonance overlap, or the need for a special NMR probe. We present here a ligand-based method that relies on the exchange broadening observed for a (13)C-labeled molecule upon binding to a protein target (labeled ligand displacement). This method can be used to screen both individual compounds and mixtures and is free of the artifacts inherent in other ligand-based methods.

[1]  Harald Schwalbe,et al.  Perspectives on NMR in drug discovery: a technique comes of age , 2008, Nature Reviews Drug Discovery.

[2]  Claudiu T. Supuran,et al.  Carbonic anhydrases: novel therapeutic applications for inhibitors and activators , 2008, Nature Reviews Drug Discovery.

[3]  M. Uesugi,et al.  [Discovering high-affinity ligands for proteins: SAR by NMR]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[4]  Jean M. Severin,et al.  Discovery and Design of Novel HSP90 Inhibitors Using Multiple Fragment‐based Design Strategies , 2007, Chemical biology & drug design.

[5]  Neal Rosen,et al.  Hsp90: a novel target for cancer therapy. , 2006, Current topics in medicinal chemistry.

[6]  P. Hajduk,et al.  Discovery of a potent inhibitor of the antiapoptotic protein Bcl-xL from NMR and parallel synthesis. , 2006, Journal of medicinal chemistry.

[7]  Andrew Emili,et al.  Navigating the Chaperone Network: An Integrative Map of Physical and Genetic Interactions Mediated by the Hsp90 Chaperone , 2005, Cell.

[8]  Gregg Siegal,et al.  TINS, target immobilized NMR screening: an efficient and sensitive method for ligand discovery. , 2005, Chemistry & biology.

[9]  D G Myszka,et al.  The ABRF-MIRG'02 study: assembly state, thermodynamic, and kinetic analysis of an enzyme/inhibitor interaction. , 2003, Journal of biomolecular techniques : JBT.

[10]  Claudio Dalvit,et al.  Fluorine-NMR experiments for high-throughput screening: theoretical aspects, practical considerations, and range of applicability. , 2003, Journal of the American Chemical Society.

[11]  J. Prestegard,et al.  A straightforward NMR-spectroscopy-based method for rapid library screening. , 2002, Angewandte Chemie.

[12]  B. Meyer,et al.  Group epitope mapping by saturation transfer difference NMR to identify segments of a ligand in direct contact with a protein receptor. , 2001, Journal of the American Chemical Society.

[13]  W. Jahnke,et al.  Spin label enhanced NMR screening. , 2001, Journal of the American Chemical Society.

[14]  Rieko Ishima,et al.  Protein dynamics from NMR , 2000, Nature Structural Biology.

[15]  M. Sundström,et al.  Identification of compounds with binding affinity to proteins via magnetization transfer from bulk water* , 2000, Journal of biomolecular NMR.

[16]  Bernd Meyer,et al.  Characterization of Ligand Binding by Saturation Transfer Difference NMR Spectroscopy. , 1999, Angewandte Chemie.

[17]  L. Pearl,et al.  Identification and Structural Characterization of the ATP/ADP-Binding Site in the Hsp90 Molecular Chaperone , 1997, Cell.

[18]  R. Meadows,et al.  Structure of Bcl-xL-Bak Peptide Complex: Recognition Between Regulators of Apoptosis , 1997, Science.

[19]  A. Gronenborn,et al.  Multidimensional heteronuclear nuclear magnetic resonance of proteins. , 1994, Methods in enzymology.