title: In Situ Reconstitution of the Adenosine A2AReceptor in Spontaneously Formed Synthetic Liposomes.
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 A2Areceptor (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.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States.
authors.forename: Roberto J
authors.initials: RJ
authors.lastname: Brea
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States.
authors.forename: Christian M
authors.initials: CM
authors.lastname: Cole
authors.affiliation: Department of Pharmacology, University of California, San Diego , La Jolla, California 92093, United States.
authors.forename: Brent R
authors.initials: BR
authors.lastname: Lyda
authors.affiliation: Department of Chemistry, University of Toronto , Mississauga, Ontario L5L 1C6, Canada.
authors.forename: Libin
authors.initials: L
authors.lastname: Ye
authors.affiliation: Department of Chemistry, University of Toronto , Mississauga, Ontario L5L 1C6, Canada.
authors.forename: R Scott
authors.initials: RS
authors.lastname: Prosser
authors.affiliation: Department of Pharmacology, University of California, San Diego , La Jolla, California 92093, United States.
authors.forename: Roger K
authors.initials: RK
authors.lastname: Sunahara
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States.
authors.forename: Neal K
authors.identifier: 0000-0002-8033-9973
authors.initials: NK
authors.lastname: Devaraj
mesh_terms: D006801:Humans; D008081:Liposomes / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D015394:Molecular Structure; D043705:Receptor, Adenosine A2A / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008081:Liposomes; D043705:Receptor, Adenosine A2A