Simultaneous assessment of membrane bilayer structure and drug insertion by ¹⁹F solid-state NMR

Fluorine-19 is an ideal nucleus for studying biological systems using NMR due to its rarity in biological environments and its favorable magnetic properties. In this work, we used a mixture of monofluorinated palmitic acids (PAs) as tracers to investigate the molecular interaction of the fluorinated drug rosuvastatin in model lipid membranes. More specifically, PAs labeled at the fourth and eighth carbon positions of their acyl chains were coincorporated in phospholipid bilayers to probe different depths of the hydrophobic core. First, the ¹⁹F chemical shift anisotropy (CSA), indicative of membrane fluidity, was simultaneously determined for fatty acids (FAs) and the fluorinated drug using either slow magic-angle spinning (MAS) 1D ¹⁹F solid-state NMR (SS-NMR) or MAS 2D ¹⁹F-¹⁹F SS-NMR with CSA recoupling. Membrane heterogeneity and selective partitioning of rosuvastatin into fluid regions could thus be evidenced. We then examined the possibility of mapping intermolecular distances in bilayers, in both the fluid and gel phases, using ¹⁹F-¹⁹F and ¹H-¹⁹F correlation experiments by SS-NMR using MAS. Spatial correlations were evidenced between the two PAs in the gel phase, while contacts between the statin and the lipids were detected in the fluid phase. This work paves the way to mapping membrane-active molecules in intact membranes, and stresses the need for new labeling strategies for this purpose.