title: Cholesterol increases kinetic, energetic, and mechanical stability of the human β2-adrenergic receptor.
abstract: The steroid cholesterol is an essential component of eukaryotic membranes, and it functionally modulates membrane proteins, including G protein-coupled receptors. To reveal insight into how cholesterol modulates G protein-coupled receptors, we have used dynamic single-molecule force spectroscopy to quantify the mechanical strength and flexibility, conformational variability, and kinetic and energetic stability of structural segments stabilizing the human β(2)-adrenergic receptor (β(2)AR) in the absence and presence of the cholesterol analog cholesteryl hemisuccinate (CHS). CHS considerably increased the kinetic, energetic, and mechanical stability of almost every structural segment at sufficient magnitude to alter the structure and functional relationship of β(2)AR. One exception was the structural core segment of β(2)AR, which establishes multiple ligand binding sites, and its properties were not significantly influenced by CHS.
authors.affiliation: Department of Biosystems Science and Engineering, Eidgenössiche Technische Hochschule Zurich, 4058 Basel, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Zocher
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.forename: Søren G F
authors.initials: SG
authors.lastname: Rasmussen
authors.forename: Brian K
authors.initials: BK
authors.lastname: Kobilka
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Müller
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D002788:Cholesterol Esters / Q000737:chemistry / Q000378:metabolism*; D004734:Energy Metabolism; D006801:Humans; D007700:Kinetics; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D055550:Protein Stability; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera; D056811:Unfolded Protein Response
chemical_list: D002788:Cholesterol Esters; D011510:Proteolipids; D018343:Receptors, Adrenergic, beta-2; D011994:Recombinant Proteins; C020485:proteoliposomes; C013440:cholesteryl succinate