Journal
NATURE CHEMISTRY
Volume 5, Issue 11, Pages 958-963Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEM.1765
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Funding
- National Institutes of Health Pathway to Independence Career Development Award [GM083155]
- National Science Foundation CAREER Award [1255250]
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [1255250] Funding Source: National Science Foundation
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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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