title: Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes.
abstract: Proteoliposome reconstitution is a standard method to stabilize purified transmembrane proteins in membranes for structural and functional assays. Here we quantified intrareconstitution heterogeneities in single proteoliposomes using fluorescence microscopy. Our results suggest that compositional heterogeneities can severely skew ensemble-average proteoliposome measurements but also enable ultraminiaturized high-content screens. We took advantage of this screening capability to map the oligomerization energy of the β2-adrenergic receptor using ∼10(9)-fold less protein than conventional assays.
authors.affiliation: 1] Department of Chemistry, University of Copenhagen, Copenhagen, Denmark. [2] Nano-Science Center, University of Copenhagen, Copenhagen, Denmark. [3] Lundbeck Foundation Center for Biomembranes in Nanomedicine, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Signe
authors.initials: S
authors.lastname: Mathiasen
authors.affiliation: 1] Department of Chemistry, University of Copenhagen, Copenhagen, Denmark. [2] Nano-Science Center, University of Copenhagen, Copenhagen, Denmark. [3] Lundbeck Foundation Center for Biomembranes in Nanomedicine, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Sune M
authors.initials: SM
authors.lastname: Christensen
authors.affiliation: 1] Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Palo Alto, California, USA. [2].
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Fung
authors.affiliation: 1] Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Palo Alto, California, USA. [2] Department of Neuroscience and Pharmacology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Søren G F
authors.initials: SG
authors.lastname: Rasmussen
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, Oregon, USA.
authors.forename: Jonathan F
authors.initials: JF
authors.lastname: Fay
authors.affiliation: 1] Department of Chemistry, University of Copenhagen, Copenhagen, Denmark. [2] Nano-Science Center, University of Copenhagen, Copenhagen, Denmark. [3] Lundbeck Foundation Center for Biomembranes in Nanomedicine, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Sune K
authors.initials: SK
authors.lastname: Jorgensen
authors.affiliation: 1] Department of Chemistry, University of Copenhagen, Copenhagen, Denmark. [2] Nano-Science Center, University of Copenhagen, Copenhagen, Denmark. [3] Lundbeck Foundation Center for Biomembranes in Nanomedicine, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Salome
authors.initials: S
authors.lastname: Veshaguri
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, Oregon, USA.
authors.forename: David L
authors.initials: DL
authors.lastname: Farrens
authors.affiliation: Department of Mathematics and Statistics, University of South Alabama, Mobile, Alabama, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Kiskowski
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Palo Alto, California, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Kobilka
authors.affiliation: 1] Department of Chemistry, University of Copenhagen, Copenhagen, Denmark. [2] Nano-Science Center, University of Copenhagen, Copenhagen, Denmark. [3] Lundbeck Foundation Center for Biomembranes in Nanomedicine, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Stamou
mesh_terms: D000465:Algorithms; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods; D011510:Proteolipids / Q000737:chemistry*; D043562:Receptors, G-Protein-Coupled / Q000032:analysis / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D011510:Proteolipids; D043562:Receptors, G-Protein-Coupled; C020485:proteoliposomes
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