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.