Nanopores have the potential to revolutionize the field of protein sequencing, but due to the biochemical complexity of polypeptide sequences, they have remained mostly theoretical. In recent work, Sauciuc et al. engineer the protein nanopore CytK to produce an electroosmotic force capable of translocating unfolded polypeptides regardless of their charge distributions, an important step toward single-file protein nanopore sequencing.