Characterization of caged compounds binding to proteins by NMR spectroscopy

Photolysable caged ligands are used to investigate protein function and activity. Here, we investigate the binding properties of caged nucleotides and their photo released products to well established but evolutionary and structurally unrelated nucleotide-binding proteins, rabbit muscle creatine kinase (RMCK) and human annexin A6 (hAnxA6), using saturation transfer difference NMR spectroscopy. We detect the binding of the caged nucleotides and discuss the general implications on interpreting data collected with photolysable caged ligands using different techniques. Strategies to avoid non-specific binding of caged compound to certain proteins are also suggested.

Research highlights
► NMR spectroscopy detects photolysable caged compounds binding to receptors.
► NMR spectroscopy highlights compromised data involving caged compounds.
► NMR controls can be performed with samples identical to other methods.
► NMR spectroscopy is a useful tool for the development of new caged compounds.