Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

Signe Mathiasen; Sune M Christensen; Juan José Fung; Søren G F Rasmussen; Jonathan F Fay; Sune K Jorgensen; Salome Veshaguri; David L Farrens; Maria Kiskowski; Brian Kobilka; Dimitrios Stamou

doi:10.1038/nmeth.3062

Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

Nat Methods. 2014 Sep;11(9):931-4. doi: 10.1038/nmeth.3062. Epub 2014 Aug 3.

Affiliations

¹ 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.
² 1] Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Palo Alto, California, USA. [2].
³ 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.
⁴ Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, Oregon, USA.
⁵ Department of Mathematics and Statistics, University of South Alabama, Mobile, Alabama, USA.
⁶ Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Palo Alto, California, USA.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Algorithms*
Image Interpretation, Computer-Assisted / methods*
Microscopy, Fluorescence / methods*
Nanotechnology / methods
Proteolipids / chemistry*
Receptors, G-Protein-Coupled / analysis
Receptors, G-Protein-Coupled / chemistry
Spectrometry, Fluorescence / methods*

Substances

Proteolipids
Receptors, G-Protein-Coupled
proteoliposomes

Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

Authors

Affiliations

Abstract

Publication types

MeSH terms

Substances

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