TY - JOUR
T1 - Sequential generation of asymmetric lipid vesicles using a pulsed-jetting method in rotational wells
AU - Gotanda, Masahide
AU - Kamiya, Koki
AU - Osaki, Toshihisa
AU - Fujii, Satoshi
AU - Misawa, Nobuo
AU - Miki, Norihisa
AU - Takeuchi, Shoji
N1 - Funding Information:
We thank Y. Nozaki for the technical support. This work was partly supported by a Grant-in-Aid for Young Scientists (A) (no. JP 15H05493 to K. K.) from the Japan Society for the Promotion of Science (JSPS); the ImPACT Program of Council for Science, Technology and Innovation (Cabinet Office, Government of Japan) ; and the Regional Innovation Strategy Support Program of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan .
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/5/15
Y1 - 2018/5/15
N2 - 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.
AB - 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.
KW - Artificial cell membrane
KW - Asymmetric lipid vesicle
KW - Lipid-protein interaction
KW - Transbilayer lipid motion
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U2 - 10.1016/j.snb.2018.01.149
DO - 10.1016/j.snb.2018.01.149
M3 - Article
AN - SCOPUS:85041382871
SN - 0925-4005
VL - 261
SP - 392
EP - 397
JO - Sensors and Actuators B: Chemical
JF - Sensors and Actuators B: Chemical
ER -