G Protein-Coupled Receptors Incorporated into Rehydrated Diblock Copolymer Vesicles Retain Functionality

Abstract

We successfully incorporated the human serotonin receptor, G protein coupled receptor (GPCR), 5-HT1AR, in micron scale polymeric giant unilamellar protein-vesicles (pGUPs). By utilizing an agarose rehydration technique for protein incorporation, the GPCR is inserted in biased orientation with the C-terminus cytosolic and N-terminus extracellular as found in the cell plasma membrane. The GPCR is fully functional within the polymeric bilayer exhibiting responses to various ligands. The entire population of incorporated GPCRs displaying activity in pGUPs remains fully functional after lyophilization for 120 hours.

Light‐Triggered Unique Shape Transformation of Giant Polymersomes with Tubular Protrusions

“…Giant polymersomes based on PB-b-PEO was prepared via agaroseassisted hydration method. [13,23] Briefly, 2% w/v agarose was dissolved in deionized water at 90 °C and spread onto a 25 mm-diameter #1 coverslip to form agarose gel. The polymer solution in chloroform (2 mg mL −1 ) with 0.5 mol% of Liss Rhod DMPE was spread on the agarose gel-coated coverslip to form a thin polymer film.…”

Durable vesicles for reconstitution of membrane proteins in biotechnology

“…Furthermore, while conventional thinking says that block copolymers for IMP reconstitution should be in a fluid state, above their glass transition temperature, proteorhodopsin (PR) has been functionally reconstituted into highly stable glassy block copolymer membranes with PS hydrophobic blocks [ 43 ]. Proteopolymersomes have also shown stability under lyophilisation: following rehydration, PBd–PEO vesicles are restored without a significant loss of IMP function, in this case a G-protein-coupled receptor [ 50 ].…”

Recent advances in permeable polymersomes: fabrication, responsiveness, and applications

“…42 (2) Adding pore-forming proteins to induce the formation of channels in the polymeric membranes which enhances the polymersome permeability and allows for selective molecule exchange. 43 (3) Utilizing stimuli-responsive constituents to endow the polymersomes with triggered-induced (such as pH, CO 2 , light, temperature, enzyme, etc. [44][45][46][47][48] ) permeability control.…”