Probing substrate binding inside a paramagnetic cavity: a NMR spectroscopy toolbox for combined experimental and theoretical investigation

Abstract

Protein cavities often rely on the paramagnetic metal present in their active site in order to catalyse various chemical transformations in biology. The selective detection and identification of the substrate is of fundamental importance in environmental monitoring and biological studies. Herein, a covalently linked Fe(III)porphyrin dimer-based paramagnetic sensory cavity has been devised for the accurate detection and simultaneous identification of phenol (substrate) binding within the cavity that provides a unique spectroscopic signature with valuable structural and environmental information. These substrates within the paramagnetic cavity leave the fingerprints of the specific binding modes (exo vs. endo) which are well distinguished with the help of various spectroscopic studies viz. UV-vis, ¹H, and ¹⁹F NMR and in their respective crystal structures also. The theoretical ¹⁹F NMR analysis plays a pivotal role in replicating the observed NMR trends with large chemical shifts of the phenolato species which in turn helps in deciphering the selective binding modes of the phenols and thereby recognizing the chemical environment within the cavity. These findings will help develop an excellent diagnostic tool for in situ monitoring of subtle conformational changes and transient interactions.