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J Am Chem Soc. 2017 Mar 15;139(10):3607-3610. doi: 10.1021/jacs.6b12830. Epub 2017 Mar 6.

In Situ Reconstitution of the Adenosine A2A Receptor in Spontaneously Formed Synthetic Liposomes.

Author information

1
Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States.
2
Department of Pharmacology, University of California, San Diego , La Jolla, California 92093, United States.
3
Department of Chemistry, University of Toronto , Mississauga, Ontario L5L 1C6, Canada.

Abstract

Cell transmembrane receptors play a key role in the detection of environmental stimuli and control of intracellular communication. G protein-coupled receptors constitute the largest transmembrane protein family involved in cell signaling. However, current methods for their functional reconstitution in biomimetic membranes remain both challenging and limited in scope. Herein, we describe the spontaneous reconstitution of adenosine A2A receptor (A2AR) during the de novo formation of synthetic liposomes via native chemical ligation. The approach takes advantage of a nonenzymatic and chemoselective method to rapidly generate A2AR embedded phospholiposomes from receptor solubilized in n-dodecyl-β-d-maltoside analogs. In situ lipid synthesis for protein reconstitution technology proceeds in the absence of dialysis and/or detergent absorbents, and A2AR assimilation into synthetic liposomes can be visualized by microscopy and probed by radio-ligand binding.

PMID:
28263576
PMCID:
PMC5702554
DOI:
10.1021/jacs.6b12830
[Indexed for MEDLINE]
Free PMC Article

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