A Dynamic NMR Lineshape Simulation Framework for Lipid Diffusion and Membrane Thinning in Bicelles and Nanodiscs

Membrane mimetics such as lipid bicelles and nanodiscs have become indispensable platforms for high-resolution structural, dynamical, and functional studies of membrane-associated systems by NMR spectroscopy, cryo-electron microscopy, and X-ray crystallography. In particular, magnetically aligned bicelles and nanodiscs uniquely enable the measurement of anisotropic NMR interactions, providing direct access to membrane geometry, lipid order, thickness, and molecular dynamics. However, the quantitative interpretation of such anisotropic NMR spectra has been hindered by the absence of physically rigorous dynamic models that properly account for the coupled effects of molecular diffusion, orientational distribution, and membrane deformation. Here, we present a comprehensive theoretical framework for the dynamic simulation of ³¹P chemical shift anisotropy and ¹⁴N quadrupolar NMR lineshapes in bicelles and nanodiscs. The model explicitly incorporates lipid diffusion, orientational distributions on curved membrane geometries, and membrane thinning, enabling physically consistent and quantitatively accurate reproduction of experimentally observed anisotropic lineshapes. Using this framework, we simulate dynamic ³¹P and ¹⁴N NMR spectra of DMPC/DHPC bicelles and nanodiscs and demonstrate how membrane thinning and lipid diffusion govern the apparent reduction of anisotropic interactions commonly observed upon peptide or protein association. This approach establishes a general physical basis for interpreting anisotropic NMR spectra of aligned membrane mimetics and provides a unified platform for quantitative investigation of membrane structure, dynamics, and membrane-active biomolecular interactions.