title: Recombinant G protein-coupled receptor expression in Saccharomyces cerevisiae for protein characterization.
abstract: G protein-coupled receptors (GPCRs) are membrane proteins that mediate signaling across the cellular membrane and facilitate cellular responses to external stimuli. Due to the critical role that GPCRs play in signal transduction, therapeutics have been developed to influence GPCR function without an extensive understanding of the receptors themselves. Closing this knowledge gap is of paramount importance to improving therapeutic efficacy and specificity, where efforts to achieve this end have focused chiefly on improving our knowledge of the structure-function relationship. The purpose of this chapter is to review methods for the heterologous expression of GPCRs in Saccharomyces cerevisiae, including whole-cell assays that enable quantitation of expression, localization, and function in vivo. In addition, we describe methods for the micellular solubilization of the human adenosine A2a receptor and for reconstitution of the receptor in liposomes that have enabled its biophysical characterization.
authors: Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans, Louisiana, USA.
authors: Kory M
authors: KM
authors: Blocker
authors: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
authors: Zachary T
authors: ZT
authors: Britton
authors: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
authors: Andrea N
authors: AN
authors: Naranjo
authors: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
authors: Patrick M
authors: PM
authors: McNeely
authors: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
authors: Carissa L
authors: CL
authors: Young
authors: Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans, Louisiana, USA; Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA. Electronic address: asr@tulane.edu.
authors: Anne S
authors: AS
authors: Robinson
mesh_terms: D000818:Animals; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins
keywords: Adenosine receptors; GPCRs; Heterologous expression; Homologous recombination; Membrane mimetic environments; Membrane proteins; Vector design