Study of molecular interactions with 13C DNP-NMR.
NMR spectroscopy is an established, versatile technique for the detection of molecular interactions, even when these interactions are weak. Signal enhancement by several orders of magnitude through dynamic nuclear polarization alleviates several practical limitations of NMR-based interaction studies. This enhanced non-equilibrium polarization contributes sensitivity for the detection of molecular interactions in a single NMR transient. We show that direct (13)C NMR ligand binding studies at natural isotopic abundance of (13)C gets feasible in this way. Resultant screens are easy to interpret and can be performed at (13)C concentrations below muM. In addition to such ligand-detected studies of molecular interaction, ligand binding can be assessed and quantified with enzymatic assays that employ hyperpolarized substrates at varying enzyme inhibitor concentrations. The physical labeling of nuclear spins by hyperpolarization thus provides the opportunity to devise fast novel in vitro experiments with low material requirement and without the need for synthetic modifications of target or ligands.
Lerche
Mathilde H
MH
Albeda Research, Gamle Carlsberg Vej 10, 2500 Valby, Denmark. mathilde.lerche@albeda.dk
Meier
Sebastian
S
Jensen
Pernille R
PR
Baumann
Herbert
H
Petersen
Bent O
BO
Karlsson
Magnus
M
Duus
Jens Ø
JØ
Ardenkjaer-Larsen
Jan H
JH
D001205:Ascorbic Acid / Q000737:chemistry; D057075:Enzyme Assays; D007887:Fabaceae / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008024:Ligands; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011485:Protein Binding; D012459:Salicylates / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry; D014510:Urease / Q000737:chemistry
D008024:Ligands; D012459:Salicylates; D012709:Serum Albumin; D014510:Urease; D001205:Ascorbic Acid