Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 261, Issue -, Pages 392-397Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2018.01.149
Keywords
Artificial cell membrane; Asymmetric lipid vesicle; Transbilayer lipid motion; Lipid-protein interaction
Funding
- Japan Society for the Promotion of Science (JSPS) [JP15H05493]
- ImPACT Program of Council for Science, Technology and Innovation (Cabinet Office, Government of Japan)
- Regional Innovation Strategy Support Program of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
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Current methods of generating asymmetric lipid vesicles produce only single types of vesicles, which poses a challenge for investigation of vesicles with different lipid leaflet combinations using a single device. Here, we describe a device for sequentially generating asymmetric lipid giant vesicles (GVs) with various combinations of asymmetric lipid leaflets. Various combinations of planar asymmetric lipid bilayers are formed by sliding and contacting the water in oil (phospholipid) (W/O) droplets in the collecting and jetting wells of our device. Next, we generate asymmetric lipid vesicles using a pulsed-jetting method. We sequentially generate three types of GVs: two asymmetric GVs containing fluorescent-conjugated phospholipids (either (i) rhodamine or (ii) BODIPY) on the outer leaflet, and (iii) a symmetric GV with phosphatidylcholine (PC) on the inner and outer leaflets. The use of asymmetric GVs with various combinations of asymmetric lipid leaflets reveals that increase in membrane phosphatidylethanolamine (PE) concentration influences cinnamycin activity and promotes the phospholipid flip-flop dynamics. This system will be useful for investigating activities of proteins or peptides on GV membranes with various combinations of lipid leaflets. (C) 2018 Elsevier B.V. All rights reserved.
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