A new NMR method for the study of ligand-protein interactions exploits the unusual lifetimes of long-lived states (LLSs). The new method provides better contrast between bound and free ligands and requires a protein-ligand ratio ca. 25 times lower than for established T(1ρ) methods, thus saving on costly proteins. The new LLS method was applied to the screening of inhibitors of urokinase-type plasminogen activator (uPA), which is a prototypical target of cancer research. With only 10 μM protein, a dissociation constant (K(D)) of 180 ± 20 nM was determined for the strong ligand (inhibitor) UK-18, which can be compared with K(D) = 157 ± 39 nM determined by the established surface plasmon resonance method.
|Data di pubblicazione:||2012|
|Titolo:||Boosting the Sensitivity of Ligand-Protein Screening by NMR of Long-Lived States|
|Rivista:||JOURNAL OF THE AMERICAN CHEMICAL SOCIETY|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1021/ja303301w|
|Appare nelle tipologie:||2.1 Articolo su rivista |