Journal: bioRxiv

Article Title: Molecular basis for cellular compartmentalization by an ancient membrane fission mechanism

doi: 10.1101/2025.11.28.690958

Figure Lengend Snippet: ( A ) AlphaFold3 structural prediction of Heimdall ESCRT-IIIA showing the canonical ESCRT-III fold of seven α-helices (top) and schematic representation of the primary sequence (bottom). ( B ) Close-up view of the predicted N-terminal α0 helix (first 15 amino acids). ( C ) Helical wheel diagram of the first 11 residues showing the asymmetric distribution of hydrophobic residues on one face of the helix. ( D ) Sequence logo illustrating conservation of the first 11 residues of ESCRT-IIIA across A-type subunits from Heimdall Asgard archaea. ( E ) Membrane remodeling assay using truncated ESCRT-IIIA lacking α0 (schematic representation of the primary sequence of ΔN-ESCRT-IIIA is shown at the top). Left: 1 µM ΔN–ESCRT-IIIA (cyan) incubated with NTs (magenta) at pH 6. Right: ΔN–ESCRT-IIIA preincubated with NTs followed by addition of ah Vps4 (5 µM) and ATP (10 mM). In both conditions, no detectable constriction or fission was observed. Scale bar, 2 µm. ( F ) AFM micrographs of E. coli extract lipid bilayers and of ΔN–ESCRT-IIIA or ESCRT-IIIA (2 µM) polymerized over the membrane. ( G ) Height quantification of ΔN–ESCRT-IIIA filaments and rings, and of ESCRT-IIIA over the lipid membranes from AFM height profiles.

Article Snippet: GUVs composed of Escherichia coli total lipid extract (Avanti Polar Lipids) were generated by electroformation as previously described ( ).

Techniques: Structural Proteomics, Sequencing, Membrane, Incubation

Journal: Biomimetics

Article Title: Synergistic Membrane Disruption of E. coli Tethered Lipid Bilayers by Antimicrobial Lipid Mixtures

doi: 10.3390/biomimetics10110739

Figure Lengend Snippet: EIS screening of potential synergies between monoglyceride and fatty acid mixtures to disrupt E. coli lipid-derived tethered bilayers. Conductance (G m , upper panel) and capacitance (C m , lower panel) signals as a function of time for E. coli lipid-derived tBLM platforms due to interaction with monoglyceride and fatty acid mixtures. Two mixture series were investigated: C 10 monoglyceride (MC) and fatty acid (CA) at ( A ) 2000 µM MC alone, ( B ) 2000 µM MC + 250 µM CA, ( C ) 2000 µM MC + 1000 µM CA, and ( D ) 2000 µM MC + 4000 µM CA; and C 12 monoglyceride (GML) and fatty acid (LA) at ( E ) 500 µM GML alone, ( F ) 500 µM GML + 31 µM LA, ( G ) 500 µM GML + 125 µM LA, and ( H ) 500 µM GML + 500 µM LA. Baseline corresponds to fabricated E. coli lipid-derived tBLM platform and arrows 1 and 2 indicate mixture addition and buffer washing steps, respectively. Graphs are representative of n = 3 independent measurements.

Article Snippet: The E. coli total lipid extract in chloroform (catalog no. 100500) was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL, USA).

Techniques: Derivative Assay

Journal: Biomimetics

Article Title: Synergistic Membrane Disruption of E. coli Tethered Lipid Bilayers by Antimicrobial Lipid Mixtures

doi: 10.3390/biomimetics10110739

Figure Lengend Snippet: EIS characterization of MC/CA mixtures to inhibit E. coli lipid-derived tethered bilayers. Conductance (G m , upper panel) and capacitance (C m , lower panel) signals as a function of time for E. coli lipid-derived tBLM platforms due to interaction with MC/CA mixtures at ( A ) 100/0 mol%, ( B ) 75/25 mol%, ( C ) 50/50 mol%, and ( D ) 25/75 mol% ratios. All mixtures were tested at 2 × CMC of the binary mixture. Baseline corresponds to fabricated E. coli lipid-derived tBLM platform and arrows 1 and 2 indicate mixture addition and buffer washing steps, respectively. ( E – H ) Corresponding Bode phase plots for each case showing ‘Baseline’ (stable signal before treatment, arrow 1), ‘Treatment’ (spectrum immediately before washing, arrow 2), and ‘Post-Wash’ (spectrum after washing), were obtained by sweeping the frequency at 3 min intervals. Graphs are representative of n = 3 independent measurements.

Article Snippet: The E. coli total lipid extract in chloroform (catalog no. 100500) was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL, USA).

Techniques: Derivative Assay

Journal: Biomimetics

Article Title: Synergistic Membrane Disruption of E. coli Tethered Lipid Bilayers by Antimicrobial Lipid Mixtures

doi: 10.3390/biomimetics10110739

Figure Lengend Snippet: EIS characterization of GML/LA mixtures to inhibit E. coli lipid-derived tethered bilayers. Conductance (G m , upper panel) and capacitance (C m , lower panel) signals as a function of time for E. coli lipid-derived tBLM platforms due to interaction with GML/LA mixtures at ( A ) 100/0 mol%, ( B ) 75/25 mol%, ( C ) 50/50 mol%, and ( D ) 25/75 mol% ratios. All mixtures were tested at 2 × CMC of the binary mixture. Baseline corresponds to fabricated E. coli lipid-derived tBLM platform and arrows 1 and 2 indicate mixture addition and buffer washing steps, respectively. ( E – H ) Corresponding Bode phase plots for each case showing ‘Baseline’ (stable signal before treatment, arrow 1), ‘Treatment’ (spectrum immediately before washing, arrow 2), and ‘Post-Wash’ (spectrum after washing), were obtained by sweeping the frequency at 3 min intervals. Graphs are representative of n = 3 independent measurements.

Article Snippet: The E. coli total lipid extract in chloroform (catalog no. 100500) was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL, USA).

Techniques: Derivative Assay

Journal: Biomimetics

Article Title: Synergistic Membrane Disruption of E. coli Tethered Lipid Bilayers by Antimicrobial Lipid Mixtures

doi: 10.3390/biomimetics10110739

Figure Lengend Snippet: Schematic comparison of MC/CA and GML/LA mixture effects on tethered E. coli lipid bilayers. Individually, MC and CA have distinct, large interaction effects on E. coli membranes that induce synergistic membrane disruption due to competing membrane morphological changes. GML and LA also exhibit distinct interaction effects but the corresponding magnitudes are smaller so synergistic membrane disruption is not observed. The mixture schematics represent the treatment step when the tethered E. coli lipid bilayers are exposed to antimicrobial lipid mixtures and illustrate the relative degree of membrane permeabilization for each mixture. The depicted packing defects indicate reversible membrane permeabilization rather than stable pore formation.

Article Snippet: The E. coli total lipid extract in chloroform (catalog no. 100500) was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL, USA).

Techniques: Comparison, Membrane, Disruption