NMR is a powerful technique that can provide detailed structural information on macromolecules and how they interact with ligands. NMR is complementary to X-ray crystallography for molecules that will not crystallize. Because NMR is typically performed in solution, the conditions can be changed to have partners and cofactors present, add potential binders in real time, and utilize mixtures. In addition, only NMR is a reliable indicator of non-binding. NMR can be used as a primary screening methodology, and is especially well-suited for fragment screening, where weak binding is expected. NMR screening is exceptionally efficient, since detection and validation are simultaneously obtained. After screening for hits, hit validation is essential; especially for difficult targets (e.g., protein-protein interactions) where authentic hits bind weakly and are buried amid the false hits.
NMR is the gold standard for sorting out binding behaviors and can immediately allow modeling of the docked ligand, which is very helpful for planning a chemistry optimization strategy. NMR can provide detailed structures of peptides and macrocycles, that is molecules that generally do not crystallize. These detailed structures guide the design of analogs that are more potent, more stable, and/or more bioavailable. The sophisticated NMR techniques needed to deal with macromolecules can be applied to smaller molecules to answer complex questions of structure or stereochemistry, or to provide the most rigorous proof of structure.
NJ Bio’s NMR services include:
- Verification of screening hits; definition of binding location(s).
- NMR screening of fragment libraries to obtain leads and Targeted Protein Degrader (TPD) ligands
- Full detailed structures of proteins, protein-ligand complexes, peptides, macrocycles, oligonucleotides, and antisense molecules.
- Advanced NMR methods to determine and verify small molecule structures.
NMR is the “Gold Standard” Method for Hit Validation.
black = only protein; red = protein + compound.
Binding is validated. Key residues are identified.
Example: Mapping of key residues (on Interleukin-2 structure) indicates binding site.
NJ Bio has a dedicated 500-MHz NMR spectrometer that is optimized for all modern 2D and 3D 1H, 13C, and 15N experiments on macromolecules.