We previously speculated that the synergistically enhanced antimicrobial activity of Magainin 2 and PGLa is related to membrane adhesion, fusion, and further membrane remodeling. Here we combined computer simulations with time-resolved in vitro fluorescence microscopy, cryoelectron microscopy, and small-angle X-ray scattering to interrogate such morphological and topological changes of vesicles at nanoscopic and microscopic length scales in real time. Coarse-grained simulations revealed formation of an elongated and bent fusion zone between vesicles in the presence of equimolar peptide mixtures. Vesicle adhesion and fusion were observed to occur within a few seconds by cryoelectron microscopy and corroborated by small-angle X-ray scattering measurements. The latter experiments indicated continued and time-extended structural remodeling for individual peptides or chemically linked peptide heterodimers but with different kinetics. Fluorescence microscopy further captured peptide-dependent adhesion, fusion, and occasional bursting of giant unilamellar vesicles a few seconds after peptide addition. The synergistic interactions between the peptides shorten the time response of vesicles and enhance membrane fusogenic and disruption properties of the equimolar mixture compared with the individual peptides.

CEITEC Central European Institute of Technology Masaryk University Brno Czech Republic; National Centre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic

CEITEC Central European Institute of Technology Masaryk University Brno Czech Republic; National Centre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic; Department of Condensed Matter Physics Faculty of Science Masaryk University Brno Czech Republic

Institute of Molecular Biosciences Biophysics Division NAWI Graz University of Graz Graz Austria; BioTechMed Graz Graz Austria

Max Planck Institute of Colloids and Interfaces Science Park Golm Potsdam Germany

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Nanoparticle induced fusion of lipid membranes

Magainin 2 and PGLa in bacterial membrane mimics IV: Membrane curvature and partitioning