Abstract
Eukaryotic subcellular membrane systems, such as the nuclear envelope or endoplasmic reticulum, present a rich array of architecturally and compositionally complex supramolecular targets that are as yet inaccessible. Here we describe layer-by-layer phospholipid membrane assembly on microfluidic droplets, a route to structures with defined compositional asymmetry and lamellarity. Starting with phospholipid-stabilized water-in-oil droplets trapped in a static droplet array, lipid monolayer deposition proceeds as oil/water-phase boundaries pass over the droplets. Unilamellar vesicles assembled layer-by-layer support functional insertion both of purified and of in situ expressed membrane proteins. Synthesis and chemical probing of asymmetric unilamellar and double-bilayer vesicles demonstrate the programmability of both membrane lamellarity and lipid-leaflet composition during assembly. The immobilized vesicle arrays are a pragmatic experimental platform for biophysical studies of membranes and their associated proteins, particularly complexes that assemble and function in multilamellar contexts in vivo.
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Acknowledgements
This work was supported by a National Institutes of Health Pathway to Independence Career Development Award (GM083155) and a National Science Foundation CAREER Award (1255250) to B.M.P.
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B.M.P. and S.M. conceived and designed the experiments, analysed the resulting data and co-authored the paper. S.M. executed all experimental work.
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Matosevic, S., Paegel, B. Layer-by-layer cell membrane assembly. Nature Chem 5, 958–963 (2013). https://doi.org/10.1038/nchem.1765
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DOI: https://doi.org/10.1038/nchem.1765
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