G protein-coupled receptors (GPCRs) are located in the plasma membrane, where they mediate the response of cells to extracellular stimuli. Despite advances in GPCR biophysical characterization, a major challenge continues to be the isolation of large quantities of membrane proteins in their functional form. We report on the purification of a GPCR, namely the human adenosine receptor A2A, using solely the short hydrocarbon chain lipid 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC, di-6:0PC). After purification, the receptor remains folded and capable of binding to ligand. Using this method of purification, the addition of cholesterol was not needed to retain receptor activity and no detergent removal step was necessary. Importantly, longer chain lipids can be  added to create a biologically relevant membanes (e.g. bilayered micelles) that can also be oriented in the presence of a magnetic field, making it an especially useful biomembrane platform for NMR studies.

Isolating a class A G protein-coupled receptor in its active conformation using short chain lipids: Doing away with detergents

245th National Meeting (2013)

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