Comparison of lipid dynamics and permeability in styrene-maleic acid and diisobutylene-maleic acid copolymer lipid nanodiscs by electron paramagnetic resonance spectroscopy.

Lipid nanoparticles formed with copolymers are a new and increasingly powerful tool for studying membrane proteins, but the extent to which these systems affect the physical properties of the membrane is not completely understood. This is critical to understanding the caveats of these new systems and screening for structural and functional artifacts that might be caused in the membrane proteins they are used to study. To better understand these potential effects, the fluid properties of dipalmitoylphosphatidylcholine lipid bilayers were examined by electron paramagnetic resonance (EPR) spectroscopy with spin-labeled reporter lipids in either liposomes or incorporated into nanoparticles with the copolymers diisobutylene-maleic acid or styrene maleic acid. Lineshape analysis at varying temperatures reveal a major change in the phase transition behavior of the lipids from a sharp melting curve in liposomes to a more gradual transition in nanoparticles. Electron spin echo envelope modulation (ESEEM) spectroscopy reveals changes in water permeability between mimetic systems, which is further supported by power-saturation measurements showing increased dequenching of spin lipids in diisobutylene-maleic acid nanoparticles compared to maleic acid nanoparticles. These results suggest that diisobutylene-maleic acid nanoparticles may have more physiological fluid properties than styrene-maleic acid nanoparticles when incorporated with saturated phospholipids.