(188dq) Isolation and Characterization of Giant Plasma Membrane Vesicles Containing a2Ar and Gas for Optical Biosensing

Giant Plasma Membrane Vesicles (GPMVs) are micron-sized unilamellar vesicles that can be rapidly isolated from most mammalian cells. Because they retain the exact membrane composition of their parent cell, GPMVs have traditionally been used to investigate phase separation of membrane proteins and lipids and lipid-lipid partitioning. However, we show here that both membrane and cytoplasmic proteins can be entrapped and released in GPMVs. To exploit this, we cotransfected Human Embryonic Kidney (HEK) cells with a CFP tagged adenosine receptor A_2AR and a YFP-tagged Ga_s protein. Using confocal microscopy, we observed GPMVs containing both fusion proteins. Attempts at selectively anchoring the GPMVs as well as Forster resonance energy transfer (FRET) are discussed herein. Altogether, GPMVs have the potential to be used as a versatile tool for reagentless optical biosensing, as they are easy to generate and cheaper to maintain than fully intact cells.