G-protein coupled receptors (GPCRs) are the largest family of transmembrane proteins involved in cell signaling. By using reductionist strategies, ligand induced signaling responses of GPCRs in vitro provide further insight into fundamental knowledge of membrane protein structure and function. Common approaches to reconstitute membrane proteins including GPCRs into membranes pose many problems such as altered protein activity and inferior biomimetic environments. Recently, our lab has developed in situ lipid synthesis for protein reconstitution technology (INSYRT), a non-enzymatic and chemoselective method to efficiently reconstitute GPCRs in phospholipid vesicles without the drawbacks of common detergent based methods. We are continuing to develop this technology for the reconstitution of additional components of the adenosine A2A receptor signaling pathway. This will allow us to characterize precise molecular events such as signaling bias and allosteric effects of ligand binding within a controlled biomimetic model setting.

Functional characterization of GPCRs in synthetic membranes using in situ lipid synthesis for protein reconstitution technology

255th National Meeting (2018)

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