High-resolution structures of the M2 channel from influenza A virus reveal dynamic pathways for proton stabilization and transduction

SignificanceThe conduction of protons through the highly restricted paths of transmembrane proteins is an essential process of living systems and an intriguing problem in modern physical chemistry. The small size of the influenza M2 proton channel makes it an ideal system for the study of proton transport across a membrane. Additionally, the M2 channel has medical relevance as an anti-flu drug target. These high-resolution structures of the channel were obtained by crystallizing the protein in a membrane-like environment and reveal networks of hydrogen-bonded waters that change with temperature and pH. The locations of these waters, in conjunction with molecular dynamics simulations that predict their hydrogen bond orientations, provide insight into the mechanism of proton stabilization and transduction within the channel.The matrix 2 (M2) protein from influenza A virus is a proton channel that uses His37 as a selectivity filter. Here we report high-resolution (1.10 Å) cryogenic crystallographic structures of the transmembrane domain of M2 at low and high pH. These structures reveal that waters within the pore form hydrogen-bonded networks or “water wires” spanning 17 Å from the channel entrance to His37. Pore-lining carbonyl groups are well situated to stabilize hydronium via second-shell interactions involving bridging water molecules. In addition, room temperature crystallographic structures indicate that water becomes increasingly fluid with increasing temperature and decreasing pH, despite the higher electrostatic field. Complementary molecular dynamics simulations reveal a collective switch of hydrogen bond orientations that can contribute to the directionality of proton flux as His37 is dynamically protonated and deprotonated in the conduction cycle