ScMreB5 binds to liposomes. (A) Sequence alignment of ScMreB5 with TmMreB and EcMreB, showing the absence of amphipathic helix at the N-terminus. Amphipathic helix of EcMreB is highlighted in red. Secondary structures are labeled on top of the alignment. (B) Sequence alignment of C-terminal region of ScMreB5 with TmMreB and EcMreB shows longer C-terminal tail enriched with positively charged residues (highlighted with blue stars). (C) Sequence alignment of ScMreB5 with TmMreB and EcMreB in the region of hydrophobic loop. The residues interacting with the membrane for TmMreB and predicted residues for ScMreB5 are highlighted in red and green, respectively. (D) Crystal structure of AMP-PNP–bound ScMreB5 (PDB accession no. 7BVY), with proposed membrane insertion loop (orange) in domain IA (green). Inset: Zoomed-in view of the loop. The N- and C-terminal ends of ScMreB5 are labeled N and C, respectively. (E) A representative 12% SDS-PAGE gel of liposome pelleting assay for comparing membrane binding of ScMreB5^WT (denoted WT), with the hydrophobic loop mutants (single mutants ScMreB5^I95A and ScMreB5^W96A and double mutant ScMreB5^{I95A, W96A}, denoted as I95A, W96A, and IWA, respectively). P and S represent the pellet and supernatant fractions of the reaction. Concentrations of liposomes of composition mimicking S. citri lipids and protein used in the assay are 1 mM and 2 µM, respectively. (F) Plot showing relative intensities of the fraction of protein in the pellet corresponding to ScMreB5^WT and hydrophobic loop mutants calculated from the SDS-PAGE gels (representative gel shown in E) from three independent experiments. The error bar denotes mean with SEM; unpaired two-tailed Student’s t test; ns, P > 0.20). Source data are available for this figure: SourceData F6.