title: Glycyrrhetic acid synergistically enhances β₂-adrenergic receptor-Gs signaling by changing the location of Gαs in lipid rafts.
abstract: Glycyrrhetic acid (GA) exerts synergistic anti-asthmatic effects via a β₂-adrenergic receptor (β₂AR)-mediated pathway. Cholesterol is an important component of the structure and function of lipid rafts, which play critical roles in the β₂AR-Gs-adenylate cyclase (AC)-mediated signaling pathway. Owing to the structural similarities between GA and cholesterol, we investigated the possibility that GA enhances β₂AR signaling by altering cholesterol distribution. Azide-terminal GA (ATGA) was synthesized and applied to human embryonic kidney 293 (HEK293) cells expressing fusion β₂AR, and the electron spin resonance (ESR) technique was utilized. GA was determined to be localized predominantly on membrane and decreased their cholesterol contents. Thus, the fluidity of the hydrophobic region increased but not the polar surface of the cell membrane. The conformations of membrane proteins were also changed. GA further changed the localization of Gαs from lipid rafts to non-raft regions, resulting the binding of β₂AR and Gαs, as well as in reduced β₂AR internalization. Co-localization of β₂AR, Gαs, and AC increased isoproterenol-induced cAMP production and cholesterol reloading attenuated this effect. A speculation wherein GA enhances beta-adrenergic activity by increasing the functional linkage between the subcomponents of the membrane β₂AR-protein kinase A (PKA) signaling pathway was proposed. The enhanced efficacy of β₂AR agonists by this novel mechanism could prevent tachyphylaxis.
authors.affiliation: College of Pharmacy, State Key Laboratory of Medicinal Chemical Biology and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Shi
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Hou
authors.forename: Jie
authors.initials: J
authors.lastname: Hou
authors.forename: Penwei
authors.initials: P
authors.lastname: Pan
authors.forename: Ze
authors.initials: Z
authors.lastname: Liu
authors.forename: Min
authors.initials: M
authors.lastname: Jiang
authors.forename: Jie
authors.initials: J
authors.lastname: Gao
authors.forename: Gang
authors.initials: G
authors.lastname: Bai
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D018927:Anti-Asthmatic Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001386:Azides / Q000737:chemistry; D002784:Cholesterol / Q000378:metabolism; D000242:Cyclic AMP / Q000096:biosynthesis; D004357:Drug Synergism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism*; D006034:Glycyrrhetinic Acid / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D008560:Membrane Fluidity / Q000187:drug effects; D021962:Membrane Microdomains / Q000187:drug effects* / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D018927:Anti-Asthmatic Agents; D001386:Azides; D018343:Receptors, Adrenergic, beta-2; D002784:Cholesterol; D000242:Cyclic AMP; D019205:GTP-Binding Protein alpha Subunits, Gs; D000262:Adenylyl Cyclases; D006034:Glycyrrhetinic Acid