<p>by Kaito Hosoda, Hidetaka Kohga, Sixian Wu, Hiroyuki Tanaka, Hideki Shigematsu, Ryoji Miyazaki, Min Fey Chek, Tomoya Tsukazaki</p> Some bacteriophages encode lysis proteins that inhibit essential bacterial processes, and elucidating their mechanisms is valuable for developing antibacterial strategies against drug-resistant pathogens. We determined the cryo-EM structure of the complex between the essential <i>Escherichia coli</i> lipid II flippase MurJ and the phage lysis protein Sgl<sup>PP7</sup>. Structural, mutational, and molecular dynamics (MD) simulation analyses revealed that Sgl<sup>PP7</sup> binds along the intramembrane cleft of MurJ and inserts its C-terminal tail into the internal cavity, stabilizing MurJ in an outward-facing conformation. Structural comparison with Lys<sup>M</sup> (Sgl<sup>M</sup>) and Sgl<sup>CJ3</sup> showed that these sequence-divergent Sgl proteins stabilize MurJ in the outward-facing conformation through distinct binding modes, consistent with the possibility of convergent evolution toward MurJ inhibition.