Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes
Signe Mathiasen1, Sune M Christensen1, Juan José Fung2
11] 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.
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.
