Mechanistic insights into CFTR function from molecular dynamics analysis of electrostatic interactions

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is an ATP-gated anion channel whose function is tightly linked to its conformational dynamics and is influenced by the composition of its membrane lipid environment. Despite high-resolution three-dimensional (3D) structures, the molecular determinants that stabilize specific CFTR conformations and enable ion conduction remain incompletely understood. Here, we performed all-atom molecular dynamics (MD) simulations of the human CFTR 3D structure in both the apo and VX-770–bound states, embedded in a heterogeneous lipid bilayer, in order to systematically analyze electrostatic interactions, linking amino acids to each other as well as to anions and membrane lipids.

We identified 558 electrostatic interactions between charged and polar amino acid side chains, which we systematically mapped across the CFTR 3D structure. They are organized into specific regions, with a subset showing high frequency and conservation across simulations, suggesting a structural role in stabilizing CFTR architecture. In contrast, more transient electrostatic interactions were detected in dynamic regions potentially linked to conformational transitions or other functional roles. Irregularities in transmembrane (TM) helices often incorporate amino acids involved in electrostatic interactions. Many basic and polar residues involved in electrostatic interactions also engaged in anion coordination, underscoring their contribution to ion conduction. In addition, some showed selective interactions with cholesterol and phosphatidylserine, revealing spatially organized lipid binding, particularly at the level of the lasso and in the vicinity of the VX-770 binding site, which may mark regions important for allosteric communication. VX-770 binding preserved the global architecture of the electrostatic interaction networks but induced subtle shifts, reinforcing specific salt bridges and enhancing anion contacts, especially around a portal displayed between TM10/TM12. This one is located opposite to the main TM4/TM6 portal, whose morphology and diameter is controlled by a fluctuating salt bridge. Regardless of whether VX-770 is present or not, two exit routes also emerged from these MD simulations. Altogether, our integrative analysis highlights how dynamic electrostatic networks, together with ion and lipid interactions support CFTR’s structural plasticity and functional modulation, offering molecular insights into potentiation mechanisms and into the specific evolution of CFTR in the ABC transporter superfamily.