2) At the reported resolution the salt bridge between Asp168 and Lys271 should be interpreted carefully. A close up of the EM densities around these residues should be provided. Although the authors have mutated Asp168 and seen a 20% reduction in hemolytic activity this is not conclusive of a salt-bridge disruption. Did mutation of Lys271 also result in the same phenotype? In addition the mutation seems to affect membrane binding as the authors mention (Figure 7 legend) and the ring attachment from the membranes (subsection “Determinants of lytic activity”). Does this imply that pores are formed but are then released? Previous AFM studies by the authors (van Pee et. Al, Nano Letters, 2016) suggest the process is irreversible. Some clarity and caution interpreting this salt bridge would improve the manuscript.