JoVE Encyclopedia of Experiments
Biological Techniques
0 views • 4:10 min • July 8th, 2025
Septins are cytoskeletal proteins comprising a GTP-binding domain flanked by terminal amino and carboxyl domains, with an inherent ability to self-assemble into filaments and higher-order structures at the inner plasma membrane for diverse cellular functions.
To study septin-membrane interactions in vitro using a biological membrane mimic, obtain a cut microcentrifuge tube. Apply ultraviolet adhesive over the uncut, flat rim and position it onto a plasma-treated, hydrophilized glass coverslip. Upon ultraviolet light exposure, the adhesive cures, creating an open reaction chamber.
Transfer monodispersed zwitterionic unilamellar lipid vesicles of desired lipid composition, supplemented with mono- and divalent cation-containing buffer, into the chamber. Cations facilitate lipid vesicles to adsorb and accumulate onto the coverslip. Gently shake the chamber, initiating vesicle rupture, and incubate.
The ruptured vesicles unfold and fuse to the coverslip via their hydrophilic head groups, forming planar lipid bilayer patches. The patches' edges trigger remaining vesicles to rupture, generating a continuous planar-supported lipid bilayer, SLB. Remove excess unadsorbed vesicles with suitab
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