biostudies-literaturePucadyil TJNIMH NIH HHSNIGMS NIH HHS517-25https://www.ebi.ac.uk/biostudies/studies/S-EPMC2905081biostudies-literatureUnknown99(2)A complete mechanistic understanding of membrane-localized processes in vesicular transport, such as membrane budding and fission, requires their reconstitution with biochemically-defined components from a biochemically-defined substrate. Supported bilayers formed by vesicle fusion represent an attractive substrate for this purpose. However, conventional supported bilayers lack a sufficient membrane reservoir to recreate membrane budding and fission events. We describe the formation of supported bilayers with excess membrane reservoir (SUPER) templates from the fusion of liposomes containing negatively charged lipids on silica beads under high-ionic-strength conditions. Using a fluorescence microscopy-based assay to monitor early and late stages of supported bilayer formation, we show thatBiophysical journalSupported bilayers with excess membrane reservoir: a template for reconstituting membrane budding and fission.PMC2905081R37 MH061345R01 MH061345R01 GM042455R01.GM42455R37.MH61345Schmid SLPucadyil TJfalseSupported bilayers with excess membrane reservoir: a template for reconstituting membrane budding and fission.A complete mechanistic understanding of membrane-localized processes in vesicular transport, such as membrane budding and fission, requires their reconstitution with biochemically-defined components from a biochemically-defined substrate. Supported bilayers formed by vesicle fusion represent an attractive substrate for this purpose. However, conventional supported bilayers lack a sufficient membrane reservoir to recreate membrane budding and fission events. We describe the formation of supported bilayers with excess membrane reservoir (SUPER) templates from the fusion of liposomes containing negatively charged lipids on silica beads under high-ionic-strength conditions. Using a fluorescence microscopy-based assay to monitor early and late stages of supported bilayer formation, we show that2010-01-01T00:00:00Z2010 Jul2025-04-04T11:31:16.492Z2019-03-27T00:32:27ZS-EPMC29050812064307010.1016/j.bpj.2010.04.036