Structural dynamics of a designed peptide pore under an external electric field.

Membrane potential is essential in biological signaling and homeostasis maintained by voltage-sensitive membrane proteins. Molecular dynamics (MD) simulations incorporating membrane potentials have been extensively used to study the structures and functions of ion channels and protein pores. They can also be beneficial in designing and characterizing artificial ion channels and pores, which will guide further amino acid sequence optimization through comparison between the predicted models and experimental data. In this study, we implemented a uniform external electric field function in the GENESIS MD simulation package to investigate the conformational dynamics of de novo-designed peptide pores. Our simulations and single-channel current recording experiments demonstrate that both charged amino acid residues in the N-terminal sequence of the peptide and the membrane potential are crucial for the structural stability and dynamics of the peptide pores. This suggests that MD simulations with an external electric field enable more accurate screening of designed proteins functioning under membrane potentials, which will ultimately contribute to a deeper understanding of voltage-sensitive membrane proteins from a bottom-up synthetic biology perspective.