Membrane free-energy landscapes derived from atomistic dynamics explain nonuniversal cholesterol-induced stiffening.

All lipid membranes have inherent morphological preferences and resist deformation. Yet adaptations in membrane shape can and do occur at multiple length scales. While this plasticity is crucial for cellular physiology, the factors controlling the morphological energetics of lipid bilayers and the dominant mechanisms of membrane remodeling remain unclear. An ongoing debate regarding the universality of the stiffening effect of cholesterol underscores the challenges facing this field, both experimentally and theoretically, even for simple lipid mixtures. On the computational side, we have argued that enhanced- sampling all-atom molecular dynamics simulations are uniquely suited for quantification of membrane conformational energetics, not only because they minimize a-priori assumptions, but also because they permit analysis of bilayers in deformed states. To showcase this approach, we examine reported inconsistencies between alternative experimental measurements of bending moduli for cholesterol-enriched membranes. Specifically, we analyze lipid bilayers with different chain saturation, and compute free-energy landscapes for curvature deformations distributed over areas from ∼5 to ∼60 nm Significance: Elucidating the energetics and mechanisms of membrane remodeling is an essential step towards understanding cell physiology. This problem is challenging, however, because membrane bending involves both large-scale and atomic-level dynamics, which are difficult to measure simultaneously. A recent controversy regarding the stiffening effect of cholesterol, which is ubiquitous in animal cells, illustrates this challenge. We show how enhanced molecular-dynamics simulations can bridge this length-scale gap and reconcile seemingly incongruent observations. This approach facilitates a conceptual connection between lipid chemistry and membrane mechanics, thereby providing a solid basis for future research on remodeling phenomena, such as in membrane trafficking or viral infection.